The landscape of MHC-presented phosphopeptides yields actionable shared tumor antigens for cancer immunotherapy across multiple HLA alleles.

BACKGROUND: Certain phosphorylated peptides are differentially presented by major histocompatibility complex (MHC) molecules on cancer cells characterized by aberrant phosphorylation. Phosphopeptides presented in complex with the human leukocyte antigen HLA-A*02:01 provide a stability advantage over their non-phosphorylated counterparts. This stability is thought to contribute to enhanced immunogenicity. Whether tumor-associated phosphopeptides presented by other common alleles exhibit immunogenicity and structural characteristics similar to those presented by A*02:01 is unclear. Therefore, we determined the identity, structural features, and immunogenicity of phosphopeptides presented by the prevalent alleles HLA-A*03:01, HLA-A*11:01, HLA-C*07:01, and HLA-C*07:02. METHODS: We isolated peptide-MHC complexes by immunoprecipitation from 11 healthy and neoplastic tissue samples using mass spectrometry, and then combined the resulting data with public immunopeptidomics data sets to assemble a curated set of phosphopeptides presented by 96 samples spanning 20 distinct healthy and neoplastic tissue types. We determined the biochemical features of selected phosphopeptides by in vitro binding assays and in silico docking, and their immunogenicity by analyzing healthy donor T cells for phosphopeptide-specific multimer binding and cytokine production. RESULTS: We identified a subset of phosphopeptides presented by HLA-A*03:01, A*11:01, C*07:01 and C*07:02 on multiple tumor types, particularly lymphomas and leukemias, but not healthy tissues. These phosphopeptides are products of genes essential to lymphoma and leukemia survival. The presented phosphopeptides generally exhibited similar or worse binding to A*03:01 than their non-phosphorylated counterparts. HLA-C*07:01 generally presented phosphopeptides but not their unmodified counterparts. Phosphopeptide binding to HLA-C*07:01 was dependent on B-pocket interactions that were absent in HLA-C*07:02. While HLA-A*02:01 and HLA-A*11:01 phosphopeptide-specific T cells could be readily detected in an autologous setting even when the non-phosphorylated peptide was co-presented, HLA-A*03:01 or HLA-C*07:01 phosphopeptides were repeatedly non-immunogenic, requiring use of allogeneic T cells to induce phosphopeptide-specific T cells. CONCLUSIONS: Phosphopeptides presented by multiple alleles that are differentially expressed on tumors constitute tumor-specific antigens that could be targeted for cancer immunotherapy, but the immunogenicity of such phosphopeptides is not a general feature. In particular, phosphopeptides presented by HLA-A*02:01 and A*11:01 exhibit consistent immunogenicity, while phosphopeptides presented by HLA-A*03:01 and C*07:01, although appropriately presented, are not immunogenic. Thus, to address an expanded patient population, phosphopeptide-targeted immunotherapies should be wary of allele-specific differences.

TCR-engaging scaffolds selectively expand antigen-specific T-cells with a favorable phenotype for adoptive cell therapy.

BACKGROUND: Adoptive cell therapy (ACT) has shown promising results for the treatment of cancer and viral infections. Successful ACT relies on ex vivo expansion of large numbers of desired T-cells with strong cytotoxic capacity and in vivo persistence, which constitutes the greatest challenge to current ACT strategies. Here, in this study, we present a novel technology for ex vivo expansion of antigen-specific T-cells; artificial antigen-presenting scaffolds (Ag-scaffolds) consisting of a dextran-polysaccharide backbone, decorated with combinations of peptide-Major Histocompatibility Complex (pMHC), cytokines and co-stimulatory molecules, enabling coordinated stimulation of antigen-specific T-cells. METHODS: The capacity of Ag-scaffolds to expand antigen-specific T-cells was explored in ex vivo cultures with peripheral blood mononuclear cells from healthy donors and patients with metastatic melanoma. The resulting T-cell products were assessed for phenotypic and functional characteristics. RESULTS: We identified an optimal Ag-scaffold for expansion of T-cells for ACT, carrying pMHC and interleukin-2 (IL-2) and IL-21, with which we efficiently expanded both virus-specific and tumor-specific CD8+ T cells from peripheral blood of healthy donors and patients, respectively. The resulting T-cell products were characterized by a high frequency of antigen-specific cells with high self-renewal capacity, low exhaustion, a multifunctional cytokine profile upon antigen-challenge and superior tumor killing capacity. This demonstrates that the coordinated stimuli provided by an optimized stoichiometry of TCR engaging (pMHC) and stimulatory (cytokine) moieties is essential to obtain desired T-cell characteristics. To generate an 'off-the-shelf' multitargeting Ag-scaffold product of relevance to patients with metastatic melanoma, we identified the 30 most frequently recognized shared HLA-A0201-restricted melanoma epitopes in a cohort of 87 patients. By combining these in an Ag-scaffold product, we were able to expand tumor-specific T-cells from 60-70% of patients with melanoma, yielding a multitargeted T-cell product with up to 25% specific and phenotypically and functionally improved T cells. CONCLUSIONS: Taken together, the Ag-scaffold represents a promising new technology for selective expansion of antigen-specific CD8+ T cells directly from blood, yielding a highly specific and functionally enhanced T-cell product for ACT.

Single CAR-T cell treatment controls disseminated ovarian cancer in a syngeneic mouse model.

BACKGROUND: Treatment of some blood cancers with T cells that express a chimeric antigen receptor (CAR) against CD19 have shown remarkable results. In contrast, CAR-T cell efficacy against solid tumors has been difficult to achieve. METHODS: To examine the potential of CAR-T cell treatments against ovarian cancers, we used the mouse ovarian cancer cell line ID8 in an intraperitoneal model that exhibits disseminated solid tumors in female C57BL/6J mice. The CAR contained a single-chain Fv from antibody 237 which recognizes a Tn-glycopeptide-antigen expressed by ID8 due to aberrant O-linked glycosylation in the absence of the transferase-dependent chaperone Cosmc. The efficacy of four Tn-dependent CARs with varying affinity to Tn antigen, and each containing CD28/CD3ζ cytoplasmic domains, were compared in vitro and in vivo in this study. RESULTS: In line with many observations about the impact of aberrant O-linked glycosylation, the ID8Cosmc knock-out (ID8Cosmc-KO) exhibited more rapid tumor progression compared with wild-type ID8. Despite the enhanced tumor growth in vivo, 237 CAR and a mutant with 30-fold higher affinity, but not CARs with lower affinity, controlled advanced ID8Cosmc-KO tumors. Tumor regression could be achieved with a single intravenous dose of the CARs, but intraperitoneal administration was even more effective. The CAR-T cells persisted over a period of months, allowing CAR-treated mice to delay tumor growth in a re-challenge setting. The most effective CARs exhibited the highest affinity for antigen. Antitumor effects observed in vivo were associated with increased numbers of T cells and macrophages, and higher levels of cleaved caspase-3, in the tumor microenvironment. Notably, the least therapeutically effective CAR mediated tonic signaling leading to antigen-independent cytokine expression and it had higher levels of the immunosuppressive cytokine interleukin10. CONCLUSION: The findings support the development of affinity-optimized CAR-T cells as a potential treatment for established ovarian cancer, with the most effective CARs mediating a distinct pattern of inflammatory cytokine release in vitro. Importantly, the most potent Tn-dependent CAR-T cells showed no evidence of toxicity in tumor-bearing mice in a syngeneic, immunocompetent system.

Effects of apatinib on quality of life and immune function in older adult patients with advanced non-small cell lung cancer / 中国基层医药

Objective:To analyze the effects of apatinib on quality of life and immune function in older adult patients with advanced non-small cell lung cancer.Methods:A total of 187 older adult patients with advanced non-small cell lung cancer admitted to Taizhou Cancer Hospital from January 2017 to January 2021 were included in this study. They were divided into the control group ( n = 93) and the observation group ( n = 94). The control group was treated with carboplatin combined with pemetrexed and the observation group was treated with apatinib based on carboplatin and pemetrexed. Sign and symptoms remission was compared between the observation and control groups. The levels of tumor markers, immune function, and quality of life score were compared between the two groups before and after treatment. Results:Total remission rate in the observation group was significantly higher than that in the control group (88.30% vs. 69.89%, χ2 = 9.59, P < 0.05). After treatment, carbohydrate antigen 125, carbohydrate antigen 50, and carcinoembryonic antigen in the observation group were (16.25 ± 5.47) μg/L, (15.23 ± 3.27) μg/L and (5.91 ± 2.66) mg/L, respectively, which were significantly lower than (21.49 ± 6.61) μg/L, (19.11 ± 3.48) μg/L and (10.14 ± 2.73) mg/L in the control group ( t = 5.91, 7.86, 10.73, all P < 0.05). The percentage of CD3 + and CD4 + cells, and the ratio of CD4 +/CD8 + cells in the observation group were (69.34 ± 8.85)%, (38.15 ± 6.52)%, (1.40 ± 0.33), respectively, which were significantly higher than (64.51 ± 8.74)%, (33.55 ± 6.33)%, (1.23 ± 0.25) in the control group ( t = -3.75, -5.36, -3.97, all P < 0.05). Quality of life score was increased in each group ( P < 0.001). The amplitude of increase in quality of life score was greater in the observation group compared with the control group ( P < 0.001). Conclusion:Apatinib can effectively reduce the level of tumor markers and improve immune function in older adult patients with advanced non-small cell lung cancer and improve quality of life.

Localized ablative immunotherapy drives de novo CD8+ T-cell responses to poorly immunogenic tumors.

BACKGROUND: Localized ablative immunotherapies hold great promise in stimulating antitumor immunity to treat metastatic and poorly immunogenic tumors. Tumor ablation is well known to release tumor antigens and danger-associated molecular patterns to stimulate T-cell immunity, but its immune stimulating effect is limited, particularly against metastatic tumors. METHODS: In this study, we combined photothermal therapy with a potent immune stimulant, N-dihydrogalactochitosan, to create a local ablative immunotherapy which we refer to as laser immunotherapy (LIT). Mice bearing B16-F10 tumors were treated with LIT when the tumors reached 0.5 cm3 and were monitored for survival, T-cell activation, and the ability to resist tumor rechallenge. RESULTS: We found that LIT stimulated a stronger and more consistent antitumor T-cell response to the immunologically 'cold' B16-F10 melanoma tumors and conferred a long-term antitumor memory on tumor rechallenge. Furthermore, we discovered that LIT generated de novo CD8+ T-cell responses that strongly correlated with animal survival and tumor rejection. CONCLUSION: In summary, our findings demonstrate that LIT enhances the activation of T cells and drives de novo antitumor T-cell responses. The data presented herein suggests that localized ablative immunotherapies have great potential to synergize with immune checkpoint therapies to enhance its efficacy, resulting in improved antitumor immunity.

Therapeutic efficacy of antibody-drug conjugates targeting GD2-positive tumors.

BACKGROUND: Both ganglioside GD2-targeted immunotherapy and antibody-drug conjugates (ADCs) have demonstrated clinical success as solid tumor therapies in recent years, yet no research has been carried out to develop anti-GD2 ADCs against solid tumors. This is the first study to analyze cytotoxic activity of clinically relevant anti-GD2 ADCs in a wide panel of cell lines with varying GD2 expression and their effects in mouse models of GD2-positive solid cancer. METHODS: Anti-GD2 ADCs were generated based on the GD2-specific antibody ch14.18 approved for the treatment of neuroblastoma and commonly used drugs monomethyl auristatin E (MMAE) or F (MMAF), conjugated via a cleavable linker by thiol-maleimide chemistry. The antibody was produced in a mammalian expression system, and its specific binding to GD2 was analyzed. Antigen-binding properties and biodistribution of the ADCs in mice were studied in comparison with the parent antibody. Cytotoxic effects of the ADCs were evaluated in a wide panel of GD2-positive and GD2-negative tumor cell lines of neuroblastoma, glioma, sarcoma, melanoma, and breast cancer. Their antitumor effects were studied in the B78-D14 melanoma and EL-4 lymphoma syngeneic mouse models. RESULTS: The ch14.18-MMAE and ch14.18-MMAF ADCs retained antigen-binding properties of the parent antibody. Direct dependence of the cytotoxic effect on the level of GD2 expression was observed in cell lines of different origin for both ADCs, with IC50 below 1 nM for the cells with high GD2 expression and no cytotoxic effect for GD2-negative cells. Within the analyzed cell lines, ch14.18-MMAF was more effective in the cells overexpressing GD2, while ch14.18-MMAE had more prominent activity in the cells expressing low GD2 levels. The ADCs had a similar biodistribution profile in the B78-D14 melanoma model compared with the parent antibody, reaching 7.7% ID/g in the tumor at 48 hours postinjection. The average tumor size in groups treated with ch14.18-MMAE or ch14.18-MMAF was 2.6 times and 3.8 times smaller, respectively, compared with the control group. Antitumor effects of the anti-GD2 ADCs were also confirmed in the EL-4 lymphoma model. CONCLUSION: These findings validate the potential of ADCs targeting ganglioside GD2 in treating multiple GD2-expressing solid tumors.

The clinical relevance of humoral immune responses to Globo H-KLH vaccine adagloxad simolenin (OBI-822)/OBI-821 and expression of Globo H in metastatic breast cancer.

An international randomized phase II trial of Globo H (GH) vaccine, adagloxad simolenin/OBI-821 in 349 patients with metastatic breast cancer showed longer progression-free survival (PFS) in vaccinated patients who developed anti-Globo H (anti-GH) IgG than those who did not and the placebo group. The impacts of anti-GH IgM and GH expression on peak anti-GH IgG and clinical outcome were further evaluated. The titers of anti-GH IgG and IgM were determined by ELISA. GH expression in tumor was examined by immunohistochemical staining. Immunophenotyping was conducted by flow cytometry. Adagloxad simolenin elicited anti-GH IgM which peaked at titers ≥1:80 between weeks 5 and 13. The mean anti-GH IgG titer peaked at week 41 and decreased thereafter on the completion of vaccination. One log increase in peak IgM was associated with 10.6% decrease in the HR of disease progression (HR: 0.894, 95% CI: 0.833 to 0.960, p=0.0019). Patients with anti-GH IgM ≥1:320 within first 4 weeks after vaccination had significantly higher maximum anti-GH IgM (p<0.0001) and IgG titers (p<0.0001) than those with <1:320. Moreover, the median PFS appears to be longer for patients with anti-GH IgM ≥1:320 within first 4 weeks than those with anti-GH IgM titer <1:320 (11.1 vs 7.3 months, p=0.164), but not statistically significant. Among patients with H score ≥80 for GH expression by immunohistochemistry, the vaccination group (n=42) seemed to have better PFS than the placebo group (n=23) (HR=0.59; 95% CI: 0.32 to 1.10, p=0.10), but the difference did not reach statistical significance. In addition, peak levels of anti-GH IgM were higher in patients who had lower percentage of activated regulatory T cells (Treg cells; CD4+CD45RA-Foxp3high) at baseline than those who had higher activated Treg cells (p=0.042). This study demonstrates that adagloxad simolenin induced both IgG and IgM antibodies against GH. Anti-GH IgM ≥1:320 within first 4 weeks or low activated Treg cells at baseline may help to select patients who are likely to produce a higher level of GH-specific IgM and IgG in the future.

The relationship between tumor volume changes, squamous cell carcinoma antigen, carcinoembryonic antigen, carbohydrate antigen 125 and the prognosis of cervical cancer patients with concurrent radiotherapy / 中国医师进修杂志

Objective:To investigate the relationship between tumor volume changes, squamous cell carcinoma antigen (SCC-Ag), carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125) and the prognosis of cervical cancer patients with concurrent radiotherapy and chemotherapy and their combined prediction of prognosis.Methods:One hundred and twenty-eight patients in Shanxi Cancer Hospital from February 2018 to February 2020, with cervical cancer undergoing radical concurrent radiotherapy and chemotherapy were selected for a prospective study. According to different prognostic effects, the patients were divided into poor prognosis group (44 cases) and good prognosis group (84 cases). The general data, tumor reduction rate (TVRR), SCC-Ag, CEA, and CA125 levels were compared between the two groups, and the Logistic regression equation was used to analyze the prognostic factors of patients with concurrent radiotherapy and chemotherapy for cervical cancer. The receiver operating characteristic (ROC) curve and the area under the curve (AUC) were used to analyze the performance of each index and the joint prediction of prognosis. Kaplan-Meier survival curve analysis and log-rank (Mantel-Cox) were used to test the survival curves of TVRR, SCC-Ag, CEA, CA125 high-risk individuals and low-risk individuals.Results:The TVRR in the poor prognosis group was significantly lower than that in the good prognosis group: (76.63 ± 7.52)% vs. (85.54 ± 6.71)%, the SCC-Ag, CEA, CA125 were significantly higher than those in the good prognosis group: (6.98 ± 2.15) μg/L vs. (4.61 ± 1.37) μg/L, (9.34 ± 2.23) μg/L vs. (5.76 ± 1.87) μg/L, (68.79 ± 12.01) kU/L vs. (49.97 ± 15.22) kU/L, and there were statistical differences ( P<0.05). Logistic regression showed that TVRR, SCC-Ag, CEA and CA125 were significant factors influencing the prognosis of patients with concurrent chemoradiotherapy for cervical cancer ( P<0.05). Among the single indicators, TVRR predicted the highest prognosis AUC, and the combined prognostic AUC of all indicators (0.837, 95% CI 0.761 to 0.920) was higher than any single indicator, with a sensitivity of 81.82% and specificity of 84.52%. The survival curves of TVRR, SCC-Ag, CEA, CA125 between high-risk and low-risk patients showed statistically significant differences ( P<0.05). Conclusions:The changes in tumor volume, SCC-Ag, CEA, CA125 and the prognosis of patients with concurrent radiotherapy and chemotherapy for cervical cancer have a certain correlation. The combined examination of the four in the early stage is expected to become a new approach to clinically predict the prognosis of cervical cancer and make appropriate treatment plans.

Clinical efficacy of GnRH-a combined with a levonorgestrel-releasing intrauterine system (Mirena) in patients with adenomyosis and its effects on ovarian function, CA125, CA153 and CEA expression / 中国基层医药

Objective:To investigate the efficacy of gonadotropin releasing hormone agonist (GnRH-a) combined with a levonorgestrel-releasing intrauterine system (Mirena) in patients with adenomyosis and its effects on ovarian function, carbohydrate antigen (CA) 125, CA153 and carcino-embryonic antigen (CEA) expression.Methods:Seventy-eight patients with adenomyosis who received treatment from September 2017 to September 2020 in Shaoxing People's Hospital were included in this study. They were randomly divided into treatment and control groups ( n = 39/group). Patients in the control group were treated with a levonorgestrel-releasing intrauterine system. Patients in the treatment group were treated with GnRH-a, once per month in the first 3 months based on treatment with a levonorgestrel-releasing intrauterine system. After 6 months of treatment, changes in dysmenorrheal relief, menstrual volume, uterine volume, endometrial thickness, ovarian function, CA125, CA153 and CEA levels relative to before treatment were compared. Results:Visual analog scale score and pictorial blood assessment chart score in the treatment group were (1.36 ± 0.28) points and (38.98 ± 5.42) points, which were significantly lower than those in the control group [(1.78 ± 0.31) points, (63.42 ± 6.75) points, t = 6.27, 17.63, both P < 0.05). Uterine volume and endometrial thickness in the treatment group were (209.74 ± 15.65) cm 3 and (7.37 ± 0.57) mm, respectively, which were significantly lower than those in the control group [(278.39 ± 20.90) cm 3, (8.63 ± 0.86) mm, t = 16.45, 7.62, P < 0.05]. There were no significant differences in serum levels of luteinizing hormone, follicle stimulating hormone and estradiol between the two groups (all P > 0.05). Serum CA125, CA153 and CEA levels in the treatment group were (26.87 ± 7.21) U/L, (23.12 ± 7.38) U/mL and (5.45 ± 0.96) μg/L, respectively, which were significantly lower than those in the control group [(49.93 ± 8.97) U/L, (38.94 ± 6.21) U/mL, (8.23 ± 1.35) μg/L, t = 12.51,10.24,10.48, P < 0.05]. Conclusion:GnRH-a combined with a levonorgestrel-releasing intrauterine system (Mirena) can markedly relieve dysmenorrhea, reduce menstrual volume, uterine volume, and endometrial thickness, has no obvious effects on ovarian function, and greatly reduce the levels of CA125, CA153 and CEA. Therefore, the combined method is a safe and effective non-surgical treatment method of adenomyosis.

Value of different tumor markers for the diagnosis and prognosis prediction of colon cancer in patients / 中国基层医药

Objective:To comprehensively analyze the clinical values of serum tumor markers of colon cancer, including carbohydrate antigen 724 (CA724), cytokeratin 19 fragment antigen (CYFRA21-1), carbohydrate antigen199 (CA199) and carcinoembryonic antigen (CEA) in the diagnosis and prognosis prediction of colon cancer in patients.Methods:The clinical data of 160 patients with colon cancer who received treatment in Zhuji Central Hospital from January 2018 to December 2020 (observation group) and the clinical data of 156 patients with benign colon polyps who concurrently received physical examination (control group) were retrospectively analyzed. All patients underwent CA724, CYFRA21-1, CA199 and CEA tumor marker screening. The levels of tumor markers, the positive rate of a single tumor marker, and the positive rate of a combination of four markers were compared between the control and observation groups. The levels of tumor markers were compared among different pathological stages. The levels of serum tumor markers were compared among patients with different prognoses based on 1-year follow-up data.Results:CA199-positve rate, CEA-positive rate, CYFRA21-1-positve rate, CA724-positive rate, and the positive rate of a combination of four tumor markers were 85.63% (137/160), 86.88% (139/160), 71.88% (115/160), 85.00% (136/160), and 95.63%(153/160), respectively, which were significantly higher than those in the control group ( χ2 = 8.64, 10.28, 8.33, 9.93, 7.27, all P < 0.001). Serum CA199, CEA, CYFRA21-1 and CA724 levels in patients with stage III-IV colon cancer were (58.96 ± 13.59) U/mL, (38.69 ± 11.84) μg/L, (14.78 ± 3.68) μg/L, (23.68 ± 5.38) U/mL, respectively, which were significantly higher than those in patients with stage I-II colon cancer [(48.35 ± 9.03) U/mL, (23.96 ± 12.25) μg/L, (9.57 ± 2.53) μg/L, (13.02 ± 4.32) U/mL, t = 10.29, 12.02, 8.47, 10.54, all P < 0.001). One-year follow-up results showed that serum levels of CA199, CEA, CYFRA21-1, CA724 in patients with recurrence and metastasis of colon cancer were (38.68 ± 3.04) U/mL, (17.12 ± 4.96) μg/L, (8.94 ± 2.32) μg/L, (11.22 ± 1.94) U/mL, which were significantly higher than those in patients without recurrence of colon cancer [(30.02 ± 2.95) U/mL, (3.75 ± 1.06) μg/L, (3.06 ± 1.15) μg/L, (6.28 ± 1.53) U/mL, t = 8.73, 11.02, 7.72, 7.57, all P < 0.001]. Conclusion:Serum levels of CEA, CA199, CA724 and CYFRA21-1 can be used as important indicators for diagnosis and prognosis prediction of colon cancer.

The value of plasma SEPT9 gene methylation combined with serum carcinoembryonic antigen and carbohydrate antigen 724 in the diagnosis of colorectal cancer / 肿瘤研究与临床

Objective:To investigate the clinical application value of plasma SEPT9 gene methylation combined with serum carcinoembryonic antigen (CEA) and carbohydrate antigen 724 (CA724) in the diagnosis of colorectal cancer.Methods:A total of 219 patients with colorectal diseases in Baoji Central Hospital and Yunnan Province New Kun Hua Hospital from May 2018 to October 2021 were selected, including 149 cases of colorectal cancer and 70 cases of colorectal polyp diagnosed by pathology. A total of 100 healthy people in the same period were selected as the healthy control group. The methylation of SEPT9 gene in plasma was measured by using real-time fluorescent polymerase chain reaction (PCR), and the levels of serum CEA and CA724 were measured by using electrochemiluminescence. The expressions of three indicators in each group were compared, and the effect of every single indicator and the combination of the three indicators on the diagnosis of colorectal cancer was analyzed by using receiver operating characteristic (ROC) curve.Results:The positive rate of SEPT9 gene methylation in colorectal cancer group (74.50%, 111/149) was higher than that in colorectal polyp group (22.86%, 16/70) and healthy control group (1.00%, 1/100), and the difference was statistically significant ( P < 0.001). The positive rate of CEA in colorectal cancer group (46.98%, 70/149) was higher than that in colorectal polyp group (40.00%, 28/70) and the healthy control group (3.00%, 3/100) and the difference was statistically significant ( P < 0.001). The positive rate of CA724 in colorectal cancer group (38.93%, 58/149) was higher than that in colorectal polyp group (32.86%, 23/70) and the healthy control group (2.00%, 2/100), and the difference was statistically significant ( P < 0.001). The area under the curve (AUC) of ROC of SEPT9 gene methylation, CEA and CA724 in the single diagnosis of colorectal cancer was 0.823 (95% CI 0.753-0.891), 0.788 (95% CI 0.725-0.852) and 0.689 (95% CI 0.624-0.754), respectively. The optimal cut-off Ct value of SEPT9 gene methylation in the diagnosis of colorectal cancer was 36.5, the sensitivity was 90.30%, and the positive predictive value was 84.68%, which were higher than those of CEA and CA724. The optimal cut-off value of CEA in the diagnosis of colorectal cancer was 8.80 ng/ml, and the specificity (77.50%) and negative predictive value (78.48%) were higher than those of SEPT9 gene methylation and CA724. The sensitivity (97.66%), positive predictive value (93.98%), negative predictive value (81.25%) and AUC (0.846, 95% CI 0.749-0.944) of the combined detection of the three indexes taking the optimal cut-off value of every single indicator were higher than those of the single indicator. Conclusions:The combined detection of plasma SEPT9 gene methylation, CEA and CA724 in the diagnosis of colorectal cancer has high sensitivity and accuracy. The three combined detection can complement each other and improve the diagnostic efficiency, which is of high clinical value for the diagnosis of colorectal cancer.

Correlation between preoperative CA19-9/GGT ratio and postoperative long-term survival of distal cholangiocarcinoma / 中华肝胆外科杂志

Objective:This research aimed to study the relationship between preoperative CA19-9/GGT ratio and postoperative long-term survival in patients with distal cholangiocarcinoma.Methods:The clinical data of 121 patients with distal cholangiocarcinoma who underwent radical pancreaticoduodenectomy (PD) at the Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2013 to December 2019 were retrospectively analyzed. The ROC curve was drawn based on the preoperative CA19-9/GGT ratio and postoperative 1-year survival. Using the best cut-off value of CA19-9/GGT ratio, the 121 patients were divided into two groups: the low ratio group (CA19-9/GGT≤0.12, n=53) and the high ratio group (CA19-9/GGT>0.12, n=68). The clinical data of the two groups were compared, and the risk factors of long-term survival were analyzed. Results:There were 72 male and 49 female patients, aged (64.9±9.2) years. When compared with the high ratio group, the low ratio group had significantly less requirement for preoperative jaundice reduction, lower CA19-9, higher GGT, better tumor differentiation, and more patients without lymph node metastasis (all P<0.05). The median follow-up time was 26 months. The 1-, 3- and 5-year survival rates of the low vs. high ratio groups were 89.4% vs. 64.7%, 64.4% vs. 14.1%, 48.7% vs. 14.1%, respectively (all P<0.001). Multivariate analysis showed that CA19-9/GGT ratio>0.12 ( RR=2.802, 95% CI: 1.494-5.256), poor differentiation ( RR=1.855, 95% CI: 1.106-3.111) and lymph node metastasis ( RR=1.891, 95% CI: 1.129-3.169) were independent risk factors for long-term survival ( P<0.05). Conclusion:The ratio of CA19-9/GGT could be used as an index to predict long-term survival of patients with distal cholangiocarcinoma after PD. The smaller the ratio, the better was the long-term prognosis.

Carbohydrate antigen 15-3 as a marker of disease severity in patients with chronic hypersensitivity pneumonitis* / Antígeno carboidrato 15-3 como marcador de gravidade da doença em pacientes com pneumonite de hipersensibilidade crônica

ABSTRACT Objective: Biomarkers associated with mucin 1, such as Krebs von den Lungen-6 and carbohydrate antigen (CA) 15-3, are increased in various interstitial lung diseases. Our aim was to determine whether CA 15-3 could be considered a biomarker of disease severity in patients with chronic hypersensitivity pneumonitis (cHP). Methods: This was a prospective observational study involving adult patients with cHP. Serum levels of CA 15-3 were measured and were correlated with variables related to disease severity and extension. HRCT scans were quantitatively analyzed using a computational platform and an image analysis tool (Computer Aided Lung Informatics for Pathology Evaluation and Rating). CA 15-3 levels were normalized by logarithmic transformation. Results: The sample comprised 41 patients. The mean age of the patients was 60.1 ± 11.6 years. The mean FVC in % of predicted was 70.3% ± 17.3%, and the median of the serum level of CA 15-3 was 48.1 U/mL. CA 15-3 levels inversely correlated with FVC in % of predicted (r = −0,30; p = 0,05), DLCO in % of predicted (r = −0,54; p < 0,01), and SpO2 at the end of a 4-min step test (r = −0,59; p < 0,01), but they directly correlated with total quantitative HRCT scores (r = 0,47; p = 0,004), especially regarding ground-glass opacities (r = 0.58; p < 0,001). Conclusions: CA 15-3 is likely to be a biomarker of disease severity of patients with cHP, particularly regarding gas exchange abnormalities.

RESUMO Objetivo: Biomarcadores associados à mucina-1, tais como Krebs von den Lungen-6 e carbohydrate antigen (CA, antígeno carboidrato) 15-3, encontram-se aumentados em diversas doenças pulmonares intersticiais. Nosso objetivo foi determinar se CA 15-3 poderia ser considerado um biomarcador de gravidade de doença em pacientes com pneumonite de hipersensibilidade crônica (PHc). Métodos: Estudo prospectivo observacional envolvendo pacientes adultos com PHc. Os níveis séricos de CA 15-3 foram medidos e correlacionados com variáveis relacionadas à gravidade e extensão da doença. As imagens de TCAR foram analisadas quantitativamente utilizando uma plataforma computacional e uma ferramenta de análise de imagem (Computer-Aided Lung Informatics for Pathology Evaluation and Rating). Os níveis de CA 15-3 foram normalizados por transformação logarítmica. Resultados: A amostra foi composta por 41 pacientes. A média de idade dos pacientes foi de 60,1 ± 11,6 anos. A média da CVF em % do previsto foi de 70,3% ± 17,3%, e a mediana do nível sérico de CA 15-3 foi de 48,1 U/mL. Os níveis de CA 15-3 se correlacionaram inversamente com CVF em % do previsto (r = −0,30; p = 0,05), DLCO em % do previsto (r = −0,54; p < 0,01) e SpO2 ao final de um teste de degrau de 4 minutos (r = −0,59; p < 0,01), mas se correlacionaram diretamente com a pontuação quantitativa total da TCAR (r = 0,47; p = 0,004), especialmente quanto a opacidades em vidro fosco (r = 0,58; p < 0,001). Conclusões: É provável que o CA 15-3 seja um biomarcador de gravidade de doença em pacientes com PHc, particularmente quanto a anormalidades nas trocas gasosas.

[Effect of iron overload on the level of carbohydrate antigen 199 in type 2 diabetes mellitus].

Objective: To investigate the effect of iron overload on tumor marker CA199 in patients with type 2 diabetes mellitus (T2DM). Methods: Between January 2017 and June 2018, three hundred and twenty-three hospitalized patients with T2DM in Department of Endocrinology, Tianjin Fourth Central Hospital were selected as diabetes group and another 100 healthy persons as control group. All the participants with major diseases and factors affecting serum ferritin and CA199 level were excluded. Serum ferritin and CA199 levels were measured by chemiluminescence assay. Patients with a ferritin level above the normal range were defined as iron overload group. According to serum ferritin level, 178 patients were divided into iron overload group and 145 into normal ferritin group in the diabetes group. The levels of CA199 were compared between iron overload group and normal ferritin group, and the correlation was analyzed. Results: The levels of serum ferritin [407.1 (346.7) µg/L vs 266.6 (201.1) µg/L in males, 184.7 (133.0) µg/L vs 77.4 (64.0) µg/L in females,both P<0.001] and CA199 [18.2 (19.3) µg/L vs 9.3 (9.7) µg/L, P<0.001] in diabetes group were significantly higher than those in control group. Serum CA199 levels in iron overload group were significantly higher than those in normal ferritin group (20.7 µg/L vs 15.0 µg/L, P<0.001; above intermediate value percent:53.9% vs 35.8%,P=0.001). CA199 was positively correlated with glycosylated hemoglobin (HbA1c) (r=0.280, P<0.001) and iron overload (r=0.210, P<0.001). Multiple linear regression analysis showed that iron overload (ß'=0.203, P=0.002) and HbA1c (ß(')'=0.202, P=0.001) had a significant effect on CA199 level. Conclusion: In T2DM patients, iron overload is an important factor affecting serum CA199 level.

[Clinical application value of γ-IFN release assay combined with CA-125 in diagnosis of active pulmonary tuberculosis].

Objective: To evaluate the diagnosis of interferon gamma release assay (IGRA) combined with tumor marker carbohydrate antigen-125 (CA-125) in active pulmonary tuberculosis (PTB). Methods: One hundred and three patients with active PTB (48 definite and 55 clinical diagnosed), 646 patients with non-PTB pulmonary disease and 60 normal controls hospitalized in Beijing Tongren Hospital, Capital Medical University between January 2014 and December 2016 were retrospectively investigated. Blood samples were collected to determine the IGRA and CA-125 level by enzyme-linked immunosorbent assay and electrochemiluminescence, respectively. The CA-125 level of patients with active PTB, non-PTB pulmonary disease and normal controls were compared. Subsequently, the best cut-off value of CA-125 for diagnosing PTB was calculated based on 60 active PTB cases and 60 normal controls. Methodological evaluation of IGRA, CA-125 and combination of these two tests (both positive) for active PTB diagnosing were performed based on 43 active PTB cases and all the non-PTB pulmonary disease cases. Results: The median values of CA-125 among definite and clinical diagnosis groups of active PTB were 55.00 (25.35, 156.90) U/ml and 81.50 (39.40, 138.00) U/ml, respectively. There was no difference between the two groups (U=1 093.00, P>0.05). And the CA-125 level of male and female PTB patients were also undifferentiated (U=1 124.00, P>0.05). There were statistically significant differences in CA-125 levels between the active PTB group and all other non-PTB groups (all P<0.001), including those who had ever closely contacted with TB patients. The area under the ROC curve constructed by CA-125 for diagnosing active PTB was 0.933. And the best cut-off value of CA-125 was 22.00 U/ml. Based on this cut-off value, the accuracy, sensitivity and specificity of CA-125 for diagnosing active PTB were 70.5% (486/689), 86.0% (37/43) and 69.5% (449/646). The accuracy, sensitivity and specificity of IGRA for diagnosing active PTB were 73.3% (480/689), 90.7% (39/43) and 68.3%(441/64). The accuracy, sensitivity and specificity of IGRA combined with CA-125 for diagnosing active PTB were 90.6% (624/689), 76.7% (33/43), 91.5% (591/646). Both of the accuracy and the false positive ratio of this combinational method (8.5%, 55/646) were significantly lower than two indexes individually used (χ(2)=94.461, 88.261, P<0.001). However, the false negative ratio was increased to 23.3% (10/43) by combinational method. Conclusion: IGRA combined with CA-125 has a certain clinical value in diagnosis of active PTB, especially when the evidences of bacterial is not available.

Clinical value of combined detection of serum CEA, CA199, CA125 in the diagnosis of lung cancer / 中国基层医药

Objective To investigate the diagnostic value of combined detection of carcinoembryonic antigen (CEA),carbohydrate antigen 199 (CA199) and carbohydrate antigen 125 (CA125) in lung cancer.Methods From July 2017 to February 2019,64 patients with lung cancer,40 cases of benign pulmonary lesions and 40 healthy people were selected by random lottery in Fuyang Hospital Affiliated to Anhui Medical University.Serum CEA,CA199 and CA125 levels of each group were analyzed.Results The levels of serum CEA,CA199 and CA125 in patients with lung cancer were significantly higher than those in patients with benign lung disease,the differences were statistically significant (t =18.355,P ＜ 0.001;t =11.757,P ＜ 0.001;t =10.754,P ＜ 0.001).The serum CA125 of lung benign disease group was significantly higher than the healthy group,and the difference was statistically significant (t =6.942,P ＜ 0.001).The positive rates of serum CEA,CA199 and CA125 in patients with lung cancer were significantly higher than those in the benign lung disease group and healthy group,the differences were statistically significant (x2 =22.110,P ＜0.001;x2 =13.321,P =0.002;x2 =15.125,P =0.001).The serum levels of CEA,CA199,and CA125 in patients with middle and advanced stage were significantly higher than those in the early stage,the differences between the advanced group and the early group were statistically significant (t =5.587,P ＜ 0.001;t =5.068,P ＜ 0.001;t =8.962,P ＜ 0.001).Those between the mid-term group and the early group had statistically significant differences (t =4.44,P ＜ 0.001;t =4.716,P ＜ 0.001;t =12.700,P ＜ 0.001).The CEA level of patients with advanced lung cancer was significantly higher than that of the intermediate patients.There was statistically significant difference in the CEA level between the advanced group and the mid-term group (t =3.860,P =0.001).Serum CA125 was the most sensitive in the diagnosis of lung cancer (60.84％),but had the lowest specificity (65.17％).CA125 compared with CA199,CEA,the difference was statistically significant (sensitivity:x2 =10.723,P ＜ 0.001specificity:x2 =4.203,P =0.023).The sensitivity of CEA + CA199 + CA125 combined detection was significantly higher than single detection,the difference was statistically significant (x2 =26.047,P ＜ 0.001),Conclusion The combined detection of serum CEA,CA125 and CA199 has high sensitivity in the diagnosis of lung cancer,which can improve the detection rate of early lung cancer and is worthy of clinical application.

Clinical application of combined detection of serum macrophage inhibitory cytokine-1, carcinoembryonic antigen and carbohydrate antigens for the diagnosis and prognosis of colorectal cancer in the elderly / 中华老年医学杂志

Objective@#To explore the application value of combined detection of serum macrophage inhibitory cytokine-1(MIC-1), carcinoembryonic antigen(CEA)and carbohydrate antigens in the diagnosis and prognosis of elderly patients with colorectal cancer.172 elderly patients with colorectal cancer were selected retrospectively as the observation group, 175 elderly patients with benign colorectal lesions were enrolled as the benign lesion group and 166 healthy elderly subjects as the control group.@*Methods@#The levels of MIC-1, CEA, carbohydrate antigens of 199(CA199), 724(CA724), 242(CA242), 125(CA125), and 50(CA50)were compared among the three groups, and the diagnostic efficacy was calculated.The difference in the above-mentioned indicators between patients with different stages and prognosis in the observation group were compared.@*Results@#Levels of MIC-1, CEA, CA199, CA242, CA724, CA125 and CA50 were higher in the observation group than in the benign lesion group and the control group, and the above indexes were higher in the benign lesion group than in the control group.The sensitivity, positive predictive value and negative predictive value of MIC-1 were the highest in single index test(68.6%, 72.8% and 84.6%, respectively). The specificity of CA199 was the highest(91.2%). The combined detections greatly improved sensitivity, positive predictive value and negative predictive value(89.0%, 79.3% and 94.1%, respectively). In the observation group, levels of MIC-1, CEA, CA199, CA242, CA724, CA125 and CA50 in the elderly colorectal cancer patients were higher in stage Ⅲ and Ⅳ than in stage Ⅰ and Ⅱ.The levels of MIC-1, CEA, CA199, CA242, CA724, CA125 and CA50 in elderly patients with recurrence and metastasis of colorectal cancer were higher than those in patients without recurrence and metastasis.@*Conclusions@#Combined detection of multiple indicators can improve the sensitivity of diagnosis, and is more conducive to the early diagnosis of elderly patients with colorectal cancer.

Clinical value of combined detection of serum CEA, CA199, CA125 in the diagnosis of lung cancer / 中国基层医药

Objective@#To investigate the diagnostic value of combined detection of carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA199) and carbohydrate antigen 125 (CA125) in lung cancer.@*Methods@#From July 2017 to February 2019, 64 patients with lung cancer, 40 cases of benign pulmonary lesions and 40 healthy people were selected by random lottery in Fuyang Hospital Affiliated to Anhui Medical University.Serum CEA, CA199 and CA125 levels of each group were analyzed.@*Results@#The levels of serum CEA, CA199 and CA125 in patients with lung cancer were significantly higher than those in patients with benign lung disease, the differences were statistically significant (t=18.355, P<0.001; t=11.757, P<0.001; t=10.754, P<0.001). The serum CA125 of lung benign disease group was significantly higher than the healthy group, and the difference was statistically significant(t=6.942, P<0.001). The positive rates of serum CEA, CA199 and CA125 in patients with lung cancer were significantly higher than those in the benign lung disease group and healthy group, the differences were statistically significant(χ2=22.110, P<0.001; χ2=13.321, P=0.002; χ2=15.125, P=0.001). The serum levels of CEA, CA199, and CA125 in patients with middle and advanced stage were significantly higher than those in the early stage, the differences between the advanced group and the early group were statistically significant (t=5.587, P<0.001; t=5.068, P<0.001; t=8.962, P<0.001). Those between the mid-term group and the early group had statistically significant differences (t=4.44, P<0.001; t=4.716, P<0.001; t=12.700, P<0.001). The CEA level of patients with advanced lung cancer was significantly higher than that of the intermediate patients.There was statistically significant difference in the CEA level between the advanced group and the mid-term group (t=3.860, P=0.001). Serum CA125 was the most sensitive in the diagnosis of lung cancer (60.84%), but had the lowest specificity (65.17%). CA125 compared with CA199, CEA, the difference was statistically significant (sensitivity: χ2=10.723, P<0.001 specificity: χ2=4.203, P=0.023). The sensitivity of CEA+ CA199+ CA125 combined detection was significantly higher than single detection, the difference was statistically significant(χ2=26.047, P<0.001),@*Conclusion@#The combined detection of serum CEA, CA125 and CA199 has high sensitivity in the diagnosis of lung cancer, which can improve the detection rate of early lung cancer and is worthy of clinical application.

Clinical application of combined detection of serum mcrophage inhibitory cytokine-1, carcinoembryonic antigen and carbohydrate antigens for the diagnosis and prognosis of colorectal cancer in the elderly / 中华老年医学杂志

Objective To explore the application value of combined detection of serum macrophage inhibitory cytokine-1 (MIC-1),carcinoembryonic antigen (CEA) and carbohydrate antigens in the diagnosis and prognosis of elderly patients with colorectal cancer.172 elderly patients with colorectal cancer were selected retrospectively as the observation group,175 elderly patients with benign colorectal lesions were enrolled as the benign lesion group and 166 healthy elderly subjects as the control group.Methods The levels of MIC-1,CEA,carbohydrate antigens of 199(CA199),724 (CA724),242 (CA242),125 (CA125),and 50 (CA50) were compared among the three groups,and the diagnostic efficacy was calculated.The difference in the above-mentioned indicators between patients with different stages and prognosis in the observation group were compared.Results Levels of MIC-1,CEA,CA199,CA242,CA724,CA125 and CA50 were higher in the observation group than in the benign lesion group and the control group,and the above indexes were higher in the benign lesion group than in the control group.The sensitivity,positive predictive value and negative predictive value of MIC-1 were the highest in single index test (68.6 ％,72.8％ and 84.6 ％,respectively).The specificity of CA199 was the highest(91.2 ％).The combined detections greatly improved sensitivity,positive predictive value and negative predictive value(89.0％,79.3％ and 94.1 ％,respectively).In the observation group,levels of MIC-1,CEA,CA199,CA242,CA724,CA125 and CA50 in the elderly colorectal cancer patients were higher in stage Ⅲ and Ⅳ than in stage Ⅰ and Ⅱ.The levels of MIC-1,CEA,CA199,CA242,CA724,CA125 and CA50 in elderly patients with recurrence and metastasis of colorectal cancer were higher than those in patients without recurrence and metastasis.Conclusions Combined detection of multiple indicators can improve the sensitivity of diagnosis,and is more conducive to the early diagnosis of elderly patients with colorectal cancer.

Changes of serum fatty acid synthase, carcinoembryonic antigen and carbohydrate antigen 72-4 in patients with gastric cancer and its clinical diagnostic value / 中国医师杂志

Objective To analyze the changes of serum fatty acid synthase (FAS),carcinoembryonic antigen (CEA) and carbohydrate antigen 72-4 (CA72-4) in patients with gastric cancer and its clinical diagnostic value.Methods Forty five patients with gastric cancer and 45 patients with benign gastric cancer treated in our hospital from January 2016 to December 2016 were enrolled in this study.Forty five healthy subjects were enrolled in this study.The levels of FAS,CEA,and CA72-4 in three groups were analyzed.Results The levels of FAS,CEA,and CA72-4 in patients with gasttric cancer [(12.73 ± 5.48) mg/L,(31.36 ± 14.51) ng/ml,and (39.32 ± 18.76) U/ml] were significantly higher than those in benign gastric cancer group [(2.26 ± 1.15) ng/L,(3.24 ± 1.47) ng/ml,and (3.75 ± 1.69) U/ml],and normal control group [(1.83 ± 0.92) mg/L,(2.71 ± 1.54) ng/ml,and (3.13 ± 1.82) U/ml] (P ＜ 0.05).FAS,CEA,and CA72-4 levels in patients with lymph node metastasis of gastric cancer [(13.58 ± 6.09) mg/L,(6.25 ± 11.54) ng/ml,and (41.31 ± 13.67) U/ml] were significantly higher than those without lymph node metastasis [(9.21 ± 5.42) mg/L,(28.38 ± 9.72) ng/ml,and (26.75 ± 11.86) U/ml] (P ＜ 0.05).The sensitivity of FAS,CEA,and CA72-4 in patients with gastric cancer was significantly lower than that in combined detection,the specificity of FAS,CEA,and CA72-4 in patients with gastric cancer was significantly higher than that in combination test (P ＜ 0.05).The sensitivity of FAS,CEA,and CA72-4 in patients with lymph node metastasis was significantly lower than that in combination test,and the specificity of FAS,CEA and CA72-4 was significantly higher than that of combined detection (P ＜ 0.05).Conclusions FAS,CEA,and CA72-4 can be used as indicators of gastric cancer and diagnosis of lymph node metastasis.Combined detection of three indexes can improve the diagnostic sensitivity and have good clinical significance.