Machine Learning and Radiomics of Bone Scintigraphy: Their Role in Predicting Recurrence of Localized or Locally Advanced Prostate Cancer.

BACKGROUND: Machine-learning (ML) and radiomics features have been utilized for survival outcome analysis in various cancers. This study aims to investigate the application of ML based on patients' clinical features and radiomics features derived from bone scintigraphy (BS) and to evaluate recurrence-free survival in local or locally advanced prostate cancer (PCa) patients after the initial treatment. METHODS: A total of 354 patients who met the eligibility criteria were analyzed and used to train the model. Clinical information and radiomics features of BS were obtained. Survival-related clinical features and radiomics features were included in the ML model training. Using the pyradiomics software, 128 radiomics features from each BS image's region of interest, validated by experts, were extracted. Four textural matrices were also calculated: GLCM, NGLDM, GLRLM, and GLSZM. Five training models (Logistic Regression, Naive Bayes, Random Forest, Support Vector Classification, and XGBoost) were applied using K-fold cross-validation. Recurrence was defined as either a rise in PSA levels, radiographic progression, or death. To assess the classifier's effectiveness, the ROC curve area and confusion matrix were employed. RESULTS: Of the 354 patients, 101 patients were categorized into the recurrence group with more advanced disease status compared to the non-recurrence group. Key clinical features including tumor stage, radical prostatectomy, initial PSA, Gleason Score primary pattern, and radiotherapy were used for model training. Random Forest (RF) was the best-performing model, with a sensitivity of 0.81, specificity of 0.87, and accuracy of 0.85. The ROC curve analysis showed that predictions from RF outperformed predictions from other ML models with a final AUC of 0.94 and a p-value of <0.001. The other models had accuracy ranges from 0.52 to 0.78 and AUC ranges from 0.67 to 0.84. CONCLUSIONS: The study showed that ML based on clinical features and radiomics features of BS improves the prediction of PCa recurrence after initial treatment. These findings highlight the added value of ML techniques for risk classification in PCa based on clinical features and radiomics features of BS.

Mechanisms of ag85a/b DNA vaccine conferred immunotherapy and recovery from Mycobacterium tuberculosis-induced injury.

Our previous research developed a novel tuberculosis (TB) DNA vaccine ag85a/b that showed a significant therapeutic effect on the mouse tuberculosis model by intramuscular injection (IM) and electroporation (EP). However, the action mechanisms between these two vaccine immunization methods remain unclear. In a previous study, 96 Mycobacterium tuberculosis (MTB) H37 Rv-infected BALB/c mice were treated with phosphate-buffered saline, 10, 50, 100, and 200 µg ag85a/b DNA vaccine delivered by IM and EP three times at 2-week intervals, respectively. In this study, peripheral blood mononuclear cells (PBMCs) from three mice in each group were isolated to extract total RNA. The gene expression profiles were analyzed using gene microarray technology to obtain differentially expressed (DE) genes. Finally, DE genes were validated by real-time reverse transcription-quantitive polymerase chain reaction and the GEO database. After MTB infection, most of the upregulated DE genes were related to the digestion and absorption of nutrients or neuroendocrine (such as Iapp, Scg2, Chga, Amy2a5), and most of the downregulated DE genes were related to cellular structural and functional proteins, especially the structure and function proteins of the alveolar epithelial cell (such as Sftpc, Sftpd, Pdpn). Most of the abnormally upregulated or downregulated DE genes in the TB model group were recovered in the 100 and 200 µg ag85a/b DNA IM groups and four DNA EP groups. The pancreatic secretion pathway downregulated and the Rap1 signal pathway upregulated had particularly significant changes during the immunotherapy of the ag85a/b DNA vaccine on the mouse TB model. The action targets and mechanisms of IM and EP are highly consistent. Tuberculosis infection causes rapid catabolism and slow anabolism in mice. For the first time, we found that the effective dose of the ag85a/b DNA vaccine immunized whether by IM or EP could significantly up-regulate immune-related pathways and recover the metabolic disorder and the injury caused by MTB.

Preventive effects of Mycobacterium tuberculosis DNA vaccines on the mouse model with latent tuberculosis infection.

Background: About a quarter of the world's population with latent tuberculosis infection (LTBI) are the main source of active tuberculosis. Bacillus Calmette Guerin (BCG) cannot effectively control LTBI individuals from developing diseases. Latency-related antigens can induce T lymphocytes of LTBI individuals to produce higher IFN-γ levels than tuberculosis patients and normal subjects. Herein, we firstly compared the effects of M. tuberculosis (MTB) ag85ab and 7 latent DNA vaccines on clearing latent MTB and preventing its activation in the mouse LTBI model. Methods: A mouse LTBI model was established, and then immunized respectively with PBS, pVAX1 vector, Vaccae vaccine, ag85ab DNA and 7 kinds of latent DNAs (including rv1733c, rv2660c, rv1813c, rv2029c, rv2628, rv2659c and rv3407) for three times. The mice with LTBI were injected with hydroprednisone to activate the latent MTB. Then, the mice were sacrificed for the bacterial count, histopathological examination, and immunological evaluation. Results: Using chemotherapy made the MTB latent in the infected mice, and then using hormone treatment reactivated the latent MTB, indicating that the mouse LTBI model was successfully established. After the mouse LTBI model was immunized with the vaccines, the lung colony-forming units (CFUs) and lesion degree of mice in all vaccines group were significantly decreased than those in the PBS group and vector group (P<0.0001, P<0.05). These vaccines could induce antigen-specific cellular immune responses. The number of IFN-γ effector T cells spots secreted by spleen lymphocytes in the ag85ab DNA group was significantly increased than those in the control groups (P<0.05). In the splenocyte culture supernatant, IFN-γ and IL-2 levels in the ag85ab, rv2029c, and rv2659c DNA groups significantly increased (P<0.05), and IL-17A levels in ag85ab and rv2659c DNA groups also significantly increased (P<0.05). Compared with the PBS and vector groups, the proportion of CD4+CD25+FOXP3+ regulatory T cells in spleen lymphocytes of ag85ab, rv2660c, rv2029c, and rv3407 DNA groups were significantly reduced (P<0.05). Conclusions: MTB ag85ab and 7 kinds of latent DNA vaccines showed immune preventive efficacies on a mouse model of LTBI, especially the rv2659c, and rv1733c DNA. Our findings will provide candidates for the development of new multi-stage vaccines against TB.

Verapamil Regulates the Macrophage Immunity to Mycobacterium tuberculosis through NF-κB Signaling.

BACKGROUND: Verapamil enhances the sensitivity of Mycobacterium tuberculosis to anti-tuberculosis (TB) drugs, promotes the macrophage anti-TB ability, and reduces drug resistance, but its mechanism is unclear. Herein, we have investigated the effect of verapamil on cytokine expression in mouse peritoneal macrophages. METHODS: Macrophages from mice infected with M. tuberculosis or S. aureus were cultured with verapamil, the cytokines were detected by enzyme-linked immunosorbent assay, and the RNA was measured with quantitative real-time polymerase chain reaction and agarose gel electrophoresis. The intracellular calcium signaling was measured by confocal microscopy. RESULTS: Significantly higher levels of NF-κB, IL-12, TNF-α, and IL-1ß were observed after TB infection. The levels of NF-κB and IL-12 increased when verapamil concentration was less than 50 µg/ml, but decreased when verapamil concentration was greater than 50µg/ml. With the increase in verapamil concentration, TNF-α and IL-1ß expressed by macrophages decreased. The L-type calcium channel transcription significantly increased in M. tuberculosis rather than S. aureus-infected macrophages. Furthermore, during bacillus Calmette-Guerin (BCG) infection, verapamil stimulated a sharp peak in calcium concentration in macrophages, while calcium concentration increased mildly and decreased smoothly over time in the absence of verapamil. CONCLUSION: Verapamil enhanced macrophage immunity via the NF-κB pathway, and its effects on cytokine expression may be achieved by its regulation of intracellular calcium signaling.

Immunotherapeutic Effects of Different Doses of Mycobacterium tuberculosis ag85a/b DNA Vaccine Delivered by Electroporation.

Background: Tuberculosis (TB) is a major global public health problem. New treatment methods on TB are urgently demanded. Methods: Ninety-six female BALB/c mice were challenged with 2×104 colony-forming units (CFUs) of MTB H37Rv through tail vein injection, then was treated with 10µg, 50µg, 100µg, and 200µg of Mycobacterium tuberculosis (MTB) ag85a/b chimeric DNA vaccine delivered by intramuscular injection (IM) and electroporation (EP), respectively. The immunotherapeutic effects were evaluated immunologically, bacteriologically, and pathologically. Results: Compared with the phosphate-buffered saline (PBS) group, the CD4+IFN-γ+ T cells% in whole blood from 200 µg DNA IM group and four DNA EP groups increased significantly (P<0.05), CD8+IFN-γ+ T cells% (in 200 µg DNA EP group), CD4+IL-4+ T cells% (50 µg DNA IM group) and CD8+IL-4+ T cells% (50 µg and 100 µg DNA IM group, 100 µg and 200 µg DNA EP group) increased significantly only in a few DNA groups (P< 0.05). The CD4+CD25+ Treg cells% decreased significantly in all DNA vaccine groups (P<0.01). Except for the 10 µg DNA IM group, the lung and spleen colony-forming units (CFUs) of the other seven DNA immunization groups decreased significantly (P<0.001, P<0.01), especially the 100 µg DNA IM group and 50 µg DNA EP group significantly reduced the pulmonary bacterial loads and lung lesions than the other DNA groups. Conclusions: An MTB ag85a/b chimeric DNA vaccine could induce Th1-type cellular immune reactions. DNA immunization by EP could improve the immunogenicity of the low-dose DNA vaccine, reduce DNA dose, and produce good immunotherapeutic effects on the mouse TB model, to provide the basis for the future human clinical trial of MTB ag85a/b chimeric DNA vaccine.

Hydronephrosis in patients with cervical cancer is an indicator of poor outcome: A nationwide population-based retrospective cohort study.

ABSTRACT: Cervical cancer is a common malignancy in women. The presence of hydronephrosis in patients with cervical cancer can be a challenging clinical problem. The appropriate management of these patients and the prediction of their outcomes are concerns among gynecologists, urologists, medical oncologists, radiation oncologists, and nephrologists. We enrolled a total of 2225 patients with cervical cancer over a 12-year period from the nationwide database of Taiwan's National Health Insurance Bureau. Among them, 445 patients had concomitant hydronephrosis. The remaining 1780 patients without hydronephrosis were randomly enrolled as a control group for the analysis of associated factors. The results indicated that the proportions of patients with hypertension, chronic kidney disease, and diabetes were significantly higher in the hydronephrosis group. The hydronephrosis group showed a higher all-cause mortality than the non-hydronephrosis group (adjusted hazard ratio 3.05, 95% confidence interval 2.24-4.15, P < .001). The rates of nephrectomy and stone disease were also significantly higher in the hydronephrosis group. A higher percentage of other cancers was also observed in the hydronephrosis group than in the non-hydronephrosis group (12.36% vs 8.99%, respectively). This study shows that cervical cancer with hydronephrosis may have a higher morbidity and mortality than cervical cancer without hydronephrosis. Other factors such as human papilloma virus vaccination, smoking, and cancer staging need to be further studied.

Chinese Traditional Medicine NiuBeiXiaoHe (NBXH) Extracts Have the Function of Antituberculosis and Immune Recovery in BALB/c Mice.

BACKGROUND: The Traditional Chinese Medicine NiuBeiXiaoHe (NBXH) is a valid antituberculosis (TB) prescription from the experience of clinical practice. However, the mechanism of NBXH extracts' immunotherapy has been poorly understood. Herein, the immunotherapeutic efficacy and the differentially expressed (DE) genes of NBXH extracts were evaluated and identified in BALB/c mice. METHODS: The total RNA was extracted from peripheral blood mononuclear cells, and the DE genes were identified by gene chip. The enrichment and signaling pathway analyses were performed using Gene Ontology (GO) and KEGG database. RESULTS: It was shown that the treatment of NBXH extracts (high dose) significantly reduced mycobacteria loads and histopathological lesions in mice infected by Mycobacterium tuberculosis and resulted in 3,454 DE upregulated genes and 3,594 downregulated DE genes. Furthermore, NBXH extracts killed mycobacteria by inhibiting the supply of necessary ingredients for their growth and proliferation. They restored the disordered immune microenvironments by up- or downregulating immune and inflammation-related pathways. CONCLUSIONS: Taken together, NBXH extracts not only efficiently decreased the mycobacteria loads but also balanced the immune disorders in mice. These new findings provide a fresh perspective for elucidating the immunotherapeutic mechanism of NBXH extracts and pointed out the direction for improving the treatment efficacy of NBXH extracts.

Effects of Mycobacterium vaccae vaccine in a mouse model of tuberculosis: protective action and differentially expressed genes.

BACKGROUND: Tuberculosis is a leading cause of death worldwide. BCG is an effective vaccine, but not widely used in many parts of the world due to a variety of issues. Mycobacterium vaccae (M. vaccae) is another vaccine used in human subjects to prevent tuberculosis. In the current study, we investigated the potential mechanisms of M. vaccae vaccination by determining differentially expressed genes in mice infected with M. tuberculosis before and after M. vaccae vaccination. METHODS: Three days after exposure to M. tuberculosis H37Rv strain (5 × 105 CFU), adult BALB/c mice randomly received either M. vaccae vaccine (22.5 µg) or vehicle via intramuscular injection (n = 8). Booster immunization was conducted 14 and 28 days after the primary immunization. Differentially expressed genes were identified by microarray followed by standard bioinformatics analysis. RESULTS: M. vaccae vaccination provided protection against M. tuberculosis infection (most prominent in the lungs). We identified 2326 upregulated and 2221 downregulated genes in vaccinated mice. These changes could be mapped to a total of 123 signaling pathways (68 upregulated and 55 downregulated). Further analysis pinpointed to the MyD88-dependent TLR signaling pathway and PI3K-Akt signaling pathway as most likely to be functional. CONCLUSIONS: M. vaccae vaccine provided good protection in mice against M. tuberculosis infection, via a highly complex set of molecular changes. Our findings may provide clue to guide development of more effective vaccine against tuberculosis.

Mannose-binding lectin 2 gene polymorphisms and their association with tuberculosis in a Chinese population.

BACKGROUND: Immune- and inflammation-related genes (IIRGs) play an important role in the pathogenesis of tuberculosis (TB). However, the relationship between IIRG polymorphisms and TB risk remains unknown. In this study, the gene polymorphisms and their association with tuberculosis were determined in a Chinese population. METHODS: We performed a case-control study involving 1016 patients with TB and 507 healthy controls of Han Chinese origin. Sixty-four single-nucleotide polymorphisms (SNPs) belonging to 18 IIRGs were genotyped by the PCR-MassArray assay, and the obtained data was analyzed with χ2-test, Bonferroni correction, and unconditional logistic regression analysis. RESULTS: We observed significant differences in the allele frequency of LTA rs2229094*C (P = 0.015), MBL2 rs2099902*C (P = 0.001), MBL2 rs930507*G (P = 0.004), MBL2 rs10824793*G (P = 0.004), and IL12RB1 rs2305740*G (P = 0.040) between the TB and healthy groups. Increased TB risk was identified in the rs930507 G/G genotype (Padjusted = 0.027) under a codominant genetic model as well as in the rs2099902 (C/T + C/C) vs T/T genotype (Padjusted = 0.020), rs930507 (C/G + G/G) vs C/C genotype (Padjusted = 0.027), and rs10824793 (G/A + G/G) vs A/A genotype (Padjusted = 0.017) under a dominant genetic model after Bonferroni correction in the analysis of the overall TB group rather than the TB subgroups. Furthermore, the rs10824793_rs7916582*GT and rs10824793_rs7916582*GC haplotypes were significantly associated with increased TB risk (P = 0.001, odds ratio [OR] = 1.421, 95% confidence interval [CI]: 1.152-1.753; and P = 0.018, OR = 1.364, 95% CI: 1.055-1.765, respectively). Moreover, the rs10824793_rs7916582*AT/AT or rs10824793_rs7916582*GT/GT diplotype showed a protective (P = 0.003, OR = 0.530, 95% CI: 0.349-0.805) or harmful (P = 0.009, OR = 1.396, 95% CI: 1.087-1.793) effect against the development of TB. CONCLUSIONS: This study indicated that MBL2 polymorphisms, haplotypes, and diplotypes were associated with TB susceptibility in the Han Chinese population. Additionally, larger sample size studies are needed to further confirm these findings in the future.

Decreased capillary density in renal cell carcinoma: Evidence from a case report with micro-computerized tomography.

RATIONALE: Conventional computerized tomography (CT) examination can differentiate renal cortical tumor from urothelial carcinoma on the basis of the highly contrast-enhanced vessels in renal cortical tumors. However, the capillary distribution of renal cell carcinoma (RCC) has been under-investigated. Here, we present a micro-CT image of tumor tissue in a patient with RCC. PATIENT CONCERNS: The patient was a 72-year-old woman with a past history of diabetes mellitus and hypertension. She did not have tumor-related symptoms. DIAGNOSIS AND INTERVENTIONS: The tumor was diagnosed using abdominal CT during her yearly routine health check. After radical nephrectomy, the tumor was subjected to pathological examination and micro-CT imaging. Pathological analysis confirmed a clear cell renal carcinoma. The capillary distribution of the tumor was significantly lesser than that of the normal cortex on micro-CT image. LESSONS: Microvessels of RCC can be detected by micro-CT. We also found that the distribution of microvessels was uneven and lower than that in the normal cortex in this case. For a more general diagnosis, more micro-CT images of RCC tumors are needed.

Comparison of Three Cellular Immunoassays to Detect Tuberculosis Infection in 876 Healthy Recruits.

The currently purified protein derivative (PPD) skin test and 2 interferon (IFN)-γ release assays (IGRAs) were usually used to detect Mycobacterium tuberculosis infection. We try to evaluate the performance of these methods to detect latent tuberculosis infection (LTBI) in this study. Each subject of the 876 recruits (19.05 ± 1.55, 17-24) underwent the PPD test, enzyme-linked immunospot (ELISPOT) assay, and chemiluminescent enzyme immunoassay (CLEIA). The prevalence of LTBI among the participants, as estimated by PPD, ELISPOT, and CLEIA, was 49.89% (437/876), 25.34% (222/876), and 28.77% (252/876), respectively. Of the participants, positive results were noted in 12.79% (112/876) for both ELISPOT and PPD, 19.52% (171/876) for both CLEIA and PPD; 9.82% (86/876) for 2 IGRAs; and 6.62% (58/876) for all 3 methods. Overall, the consistency among the 3 tests was 36.99% (324/876). ELISPOT-positive rate (41.38%) in the recruits with a PPD result ≥20 mm was higher than PPD <20 mm (24.76%; P < 0.05). Increased PPD skin reactions were associated with significantly increased CLEIA-positive rates and IFN-γ levels. Of 307 recruits without the bacillus Calmette-Guérin (BCG) vaccination, 2 IGRA (42.19%)-positive rates in the PPD-positive group were significantly higher than those in the PPD-negative group (28.40% and 23.05%; P < 0.05 and P < 0.01, respectively).There was low correlation and poor consistency among 2 IGRAs and PPD in healthy recruits, but IGRAs may be more accurate screening methods for TB infection in the countries with BCG vaccination.

Immunogenicity and Therapeutic Effects of Latency-Associated Genes in a Mycobacterium Tuberculosis Reactivation Mouse Model.

In this study, the Mycobacterium tuberculosis (MTB) latency-associated antigens Rv2660c, Rv1733c, Rv1813c, Rv2628, Rv2029c, and Rv2659c were compared regarding their immunogenicity and potential therapeutic effects in an MTB reactivation mouse model. Normal mice or MTB reactivation mice were immunized intramuscularly three times at 2-week intervals with saline, plasmid vector pVAX1, Mycobacterium vaccae vaccine (a commercial inactivated vaccine), rv1813c DNA, rv2628 DNA, rv2029c DNA, rv2659c DNA, rv1733c DNA, or rv2660c DNA. The normal mice immunized with rv2628 DNA or rv2659c DNA had low numbers of Th1 cells and a lower ratio of Th1:Th2 immune cells in whole blood (p < 0.05). Compared to the saline group, Tc1 cells in the rv2029c DNA group and Tc1:Tc2 cell ratio in the rv1813c DNA, rv2628 DNA, and rv2029c DNA groups were significantly decreased (p < 0.05). The proportion of Foxp3+CD4+ T cells in the rv2628 DNA and rv2659c DNA groups and the proportion of CD4+CD25+ T cells in the rv2029c DNA group were significantly increased (p < 0.05). The level of anti-Rv1813c-immunoglobulin G (IgG) in the rv1813c DNA group was significantly increased (p < 0.01). The levels of specific IgG, IgG1, and IgG2a in the rv2628 DNA, rv2029c DNA, and rv2659c DNA groups were significantly increased (p < 0.05). Lung colony-forming units in M. vaccae and the six DNA groups decreased to different degrees in the MTB reactivation mouse model, but only the lung colony-forming units in the rv2628 DNA group (4.38 ± 0.70 log10) significantly decreased compared to the vector group (5.90 ± 0.42 log10; p < 0.05). The MTB rv1813c DNA, rv2628 DNA, rv2029c DNA, and rv2659c DNA could elicit a strong humoral immune response and a higher proportion of CD4+CD25+or CD4+Foxp3+ T cells but could not increase the proportions of Th1 and Tc1 cells. These results suggest that latency-associated DNA vaccines, especially rv2628 DNA, had some therapeutic effect on the endogenous resurgence mouse tuberculosis model.

Salvia miltiorrhiza Bunge (Danshen) for Treatment and Prevention of Urolithiasis: A Drosophila Animal Study.

Traditional Chinese medicine (TCM) has been prescribed for the treatment of stone disease for thousands of years. Salvia miltiorrhiza (Danshen) was previously shown to have potential for treatment of stone disease in animal and clinical studies. In this study, we further studied the antiurolithiasis effect of Danshen in a fly model. Wild-type male Drosophila melanogaster CS flies were used in this study, with 0.25% ethylene glycol (EG) as a lithogenic agent. 2% potassium citrate (K-citrate) was the positive control agent for prevention (all agents added at the start of experiment) and treatment (drugs added after 2-week addition of lithogenic agent) studies compared with 15, 30, and 60 µg/ml of Danshen extract. In the prevention study, both 2% K-citrate and Danshen (30 and 60 µg/ml) significantly inhibited EG-induced calcium oxalate (CaOx) crystal formation. In the treatment study, only 2% K-citrate and high-dose of Danshen (60 µg/ml) significantly inhibited EG-induced CaOx crystal formation. Survival analysis for EG with Danshen was compared with that for EG with K-citrate. The mean lifespan was significantly reduced by administration of EG, and the results in the Danshen group were similar to those in the control group. In conclusion, Danshen revealed both preventive and treatment effects on CaOx crystal formation in a fly model.

Novel epitopes identified from Mycobacterium tuberculosis antigen Rv2629induces cytotoxic T lymphocyte response.

There is an urgent need for a more effective vaccine against tuberculosis (TB). Cytotoxic T lymphocytes (CTLs) play a critical role in combating Mycobacterium tuberculosis (M.tb). The identification of novel CTL epitopes is essential for the design of peptide-based vaccines. In this study, we predicted CTL epitope peptides of M.tb antigen Rv2629 restricted by HLA-A2, using bioinformatics methods. The affinity and stability of binding of these peptides with HLA-A2 molecules were detected by flow cytometry. Their ability to induce CTLs generation was determined in peripheral blood mononuclear cells (PBMCs) from healthy uninfected subjects, Latent tuberculosis infection (LTBI) subjects, and TB patients ex vivo. The cytotoxic activity induced by the epitope peptides was tested by lactate dehydrogenase (LDH) release assay. Finally, we found four novel CTL epitope peptides, Rv2629-p190-2L, Rv2629-p190-1Y2L, Rv2629-p274, and Rv2629-p315, which had high-affinity and stability of binding with T2 cells. Their ability of inducing CTLs was highest in PBMCs from TB patients (P < 0.05). In addition, these peptides could induce the CTLs to generate specific cytotoxic activity. They showed higher immunogenicity in TB patients and had the potential to become candidate vaccines for TB therapy.

Diagnostic performance and problem analysis of commercial tuberculosis antibody detection kits in China.

BACKGROUND: The diagnosis of bacterium-negative pulmonary tuberculosis (TB) and extra-pulmonary TB is challenging clinically. The detection of the anti-TB antibody has an important, auxiliary, clinical diagnostic value. Therefore, TB antibody detection kits should be screened and evaluated, and the reagents with the highest sensitivity and specificity should be chosen and used clinically. METHODS: The diagnostic performance of 7 commercially available TB antibody detection kits (kits A, B, C, D, E, F and G) based on the gold immunoassay detection of immunoglobulin (Ig) G or IgM antibodies were simultaneously evaluated and compared in 62 TB cases and 56 non-TB cases in a laboratory. A retrospective analysis including 2549 cases was carried out to assess the clinical diagnosis values of bacteriological examinations and TB antibody tests (kits B and H used in the clinic). RESULTS: The sensitivities of TB antibody kits A, B, C, D, E, F and G in the sera from 62 TB patients were 50.0%, 83.9%, 38.7%, 9.7%, 48.4%, 69.4% and 79.0%, respectively; the sensitivities in the sera from 24 smear-negative TB patients were 29.2%, 79.2%, 29.2%, 12.5%, 29.2%, 54.2% and 79.2%, respectively; the specificities in the sera from 56 non-TB patients were 73.2%, 25.0%, 85.7%, 96.4%, 78.6%, 78.6% and 50.0%, respectively. Of the 2549 clinically diagnosed cases, there were 1752 pulmonary TB cases, 505 extra-pulmonary TB cases, 87 old pulmonary TB cases and 205 non-TB cases. The positive results for smear, culture, TB antibody kit B and kit H in pulmonary TB cases were 39.8% (543/1365), 48.6% (372/765), 45.8% (802/1752) and 25.2% (442/1752), respectively; the results in extra-pulmonary TB cases were 3.4% (6/178), 5.8% (4/69), 35.4% (179/505), and 11.3% (57/505), respectively; the results in old pulmonary TB cases were 0% (0/64), 0% (0/30), 32.2% (28/87), and 9.2% (8/87), respectively; and the results in non-TB cases were 0% (0/121), 0% (0/56), 21.5% (44/205), and 2.4% (5/205), respectively. Of 624 smear-positive and/or culture-positive pulmonary TB cases, the sensitivities of antibody test kits B and H were 53.0% and 36.4%, respectively. Of 901 smear-negative and/or culture-negative pulmonary TB cases, the sensitivities of antibody test kits B and H were 42.5% and 19.0%, respectively. The positive rate of antibody detection in the bacterium-positive pulmonary TB cases was significantly higher than that in the bacterium-negative pulmonary TB cases (P < 0.05). CONCLUSIONS: The colloidal gold-labeled TB antibody IgG detection assay is a simple, rapid and economical method that provides a better clinical auxiliary diagnosis value on TB, especially in smear-negative pulmonary TB and extra-pulmonary TB. The production, quality control, screening and evaluation of antibody detection kits are very important for its clinical application.

Cytokine and soluble adhesion molecule profiles and biomarkers for treatment monitoring in Re-treated smear-positive patients with pulmonary tuberculosis.

Relapse of pulmonary tuberculosis (PTB) is associated with a failure of the host immune system to control the invading Mycobacterium tuberculosis. Severe immunodeficiency or immune disorders may be the main reason for TB recurrence. This study aimed to quantify serum inflammatory cytokine and soluble adhesion molecule levels in Re-treated smear-positive PTB patients before and after re-anti-TB drug therapy. Serum samples were collected from 30 healthy controls and 215 Treated active PTB patients at baseline and 2, 4, and 6 months post-re-treatment. Levels of 18 serum cytokines and soluble adhesion molecules were measured by a high-throughput Cytometric Bead Array. At baseline, IL-1, IL-2, IL-12P70, and soluble CD62E levels were significantly higher in PTB patients than those in the healthy controls (p < 0.05); IL-4, IL-5, IL-7, IL-8, IL-10, IL-17, IL-21, soluble CD54, MIG, and TGF-ß levels in PTB patients were significantly lower than those in the healthy controls (p < 0.05), of which TGF-ß, IL-7, IL-8, IL-10, soluble CD54, and MIG were most notably (p < 0.0005). After re-treatment, IFN-γ, IL-2, IL-7, and soluble CD54 levels and IL-2/IL-10 and IFN-γ/IL-10 ratios showed an upward trend during the re-treatment period. They were more sensitive than other cytokines and adhesion molecules and could be effective as serum indicators for re-treatment response. The immune response was imbalance in treated smear-positive PTB patients: Th1 response was elevated, but Th2 and Th17 responses were reduced. Systematic and comprehensive understanding of the cytokine and soluble adhesion molecule profiles provides a theoretical basis for immuno-diagnosis, immunotherapy, and immuno-monitoring of Re-treated PTB patients.

Immunotherapeutic effects of Mycobacterium tuberculosis rv3407 DNA vaccine in mice.

Tuberculosis (TB) is a major global public health problem. Latent TB infection (LTBI) is a major source of active TB. New vaccines to treat LTBI are urgently demanded. In this study, the gene encoding latency-associated antigen Rv3407 of Mycobacterium tuberculosis (MTB) rv3407 DNA vaccine was used to prepare and the immunogenicity and therapeutic effects were evaluated. Normal mice were immunized intramuscularly three times at two-week intervals with sterile water for injection, plasmid vector pVAX1, M. vaccae vaccine, ag85a DNA or rv3407 DNA. TB-infected mice were immunized intramuscularly three times at two-week intervals with phosphate-buffered saline (PBS) and rv3407 DNA. The normal mice immunized with rv3407 DNA or ag85a DNA showed higher levels of interferon-gamma (IFN-γ) in stimulated spleen lymphocyte culture supernatants, and had more Th1 cells and an elevated ratio of Th1/Th2 immune cells in whole blood, indicating that a Th1-type immune response was predominant. The levels of anti-Ag85A or anti-Rv3407 IgG antibody were significantly increased in the ag85a DNA and rv3407 DNA groups compared to the sterile water for injection, vector, and M. vaccae groups (p < .0001). Compared with the PBS group, the rv3407 DNA group had pulmonary bacterial loads that were lower by 0.56 log10 (p < .01). The mice vaccinated with rv3407 DNA developed antigen-specific cellular and humoral responses. The rv3407 DNA is a potential DNA vaccine candidate against TB.

Immunogenicity and therapeutic effects of recombinant Ag85AB fusion protein vaccines in mice infected with Mycobacterium tuberculosis.

The immune function of tuberculosis (TB) patients is disordered. By using immune regulators to assist chemotherapy for TB the curative effect might be improved. In this study, a vaccine containing Mycobacterium tuberculosis (M. tuberculosis) recombinant Ag85AB fusion protein (rAg85AB) was constructed and evaluated. The mice were immunized intramuscularly three times at two-week intervals with Ag85AB fusion protein combined with Corynebacterium parvum adjuvant (rAg85AB+CP). In comparison to control mice that received either CP alone or saline, the mice that received rAg85AB+CP had significantly higher number of T cells secreting IFN-γ and higher levels of specific antibodies of IgG, IgG1 and IgG2a isotypes in sera. The specific antibodies also had higher ratios of IgG2a to IgG1, indicating a predominant Th1 immune response. To test for immunotherapy of TB, M. tuberculosis infected mice were given three intramuscular doses of 20µg, 40µg or 60µg of rAg85AB in rAg85AB+CP, or phosphate-buffered saline (PBS), or CP or Mycobacterium phlei (M. Phlei) F.U.36. Compared with the PBS group, 20µg, 40µg and 60µg rAg85AB+CP and M. phlei F.U.36 groups reduced the pulmonary bacterial loads by 0.13, 0.15, 0.42 and 0.40 log10, and the liver bacterial loads by 0.64, 0.64, 0.53 and 0.61 log10, respectively. Pathological changes of lungs were less, and the lesions were limited to a certain extent in 40µg and 60µg rAg85AB+CP and M. phlei F.U.36 groups. These results showed that rAg85AB+CP had immunotherapeutic effect on TB, significantly increasing the cellular immune response, and inhibiting the growth of M. tuberculosis.

An alert of Mycobacterium tuberculosis infection of rhesus macaques in a wild zoo in China.

Mycobacterium tuberculosis, the pathogen that causes tuberculosis (TB), is becoming increasingly recognized as an important cause of fatal chronic illnesses in China. In this study, we report an infectious disease among 84 rhesus macaques at a Chinese zoo. Their clinical signs and symptoms were very similar with the manifestations of TB in humans. To determine the potential pathogens of this outbreak, many methods were used. First, tuberculin skin tests showed that none of the monkeys displayed significant skin reactions. Subsequently, the sera were tested for specific antibody IgG; 29 (34.5%) and 39 (46.4%) blood samples tested positive by TB-IgG and TB-DOT, respectively. Radiographic examination showed characteristic imageology changes in 14 (16.7%) monkeys. One individual determined as positive by the above three methods was euthanized, and histopathological analysis demonstrated typical granulomas and caseous necrosis in the lung, liver, spleen, and intestine. Furthermore, the pathogenic mycobacteria were isolated from lung lobe, cultured on acidic Lowenstein-Jensen culture medium, and identified as M. tuberculosis by real-time PCR and DNA sequencing. Nevertheless, the origin of the infection remained unknown. These findings emphasize the need to strengthen the management and training of staff, especially those working at animal shelters.

A new method of screening for latent tuberculosis infection: Results from army recruits in Beijing in 2014.

Latent tuberculosis infection (LTBI) lacks diagnostic gold method. Effective method is important to the control of tuberculosis. IFN-γ responses in 600 military recruits were tested by ELISA using whole blood incubation with latent protein Rv2029c, Rv2659c and recombinant protein CFP10-ESAT6 (rCE) respectively. They also received tuberculin skin test. Their BCG vaccination status was recorded. When 30.7% (184/600) of recruits gave higher IFN-γ responses (≥470pg/mL) to rCE as LTBI, the rests as healthy control, the AUC of rRv2029c was 0.856 and rRv2659c was 0.827 for LTBI diagnosis. IFN-γ responses to rCE were higher in PPD-positive group (≥5mm) than negative group (<5mm) (p<0.05), while for rRv2029c and rRv2659c were not (p>0.05). IFN-γ responses induced by rRv2029c and rRv2659c were higher in the moderately-positive group (≥5, <15mm) than the strongly-positive group (≥15mm) (p<0.05), while for rCE were not (p>0.05). IFN-γ levels to three antigens were not related to BCG vaccination status (p>0.05). Rv2659c and Rv2029c are good candidate antigens to complement the role of rCE for LTBI diagnosis, which provide a basis for developing cost-effective LTBI screening methods in the army.