Correlation Between ImageJ and Conventional Manual Scoring Methods for Programmed Death-Ligand 1 Immuno-Histochemically Stained Sections.

Background: One of the most frequently used methods for quantifying PD-L1 (programmed cell death-ligand 1) expression in tumor tissue is IHC (immunohistochemistry). This may predict the patient's response to anti-PD1/PD-L1 therapy in cancer. Methods: ImageJ software was used to score IHC-stained sections for PD-L1 and compare the results with the conventional manual method. Results: In diffuse large B cell lymphoma, no significant difference between the scores obtained by the conventional method and ImageJ scores obtained using the option "RGB" or "Brightness/Contrast." On the other hand, a significant difference was found between the conventional and HSB scoring methods. ImageJ faced some challenges in analyzing head and neck squamous cell carcinoma tissues because of tissue heterogenicity. A significant difference was found between the conventional and ImageJ scores using HSB or RGB but not with the "Brightness/Contrast" option. Scores obtained by ImageJ analysis after taking images using 20 × objective lens gave significantly higher readings compared to 40 × magnification. A significant difference between camera-captured images' scores and scanner whole slide images' scores was observed. Conclusion: ImageJ can be used to score homogeneous tissues. In the case of highly heterogeneous tissues, it is advised to use the conventional method rather than ImageJ scoring.

COVID-19 Antibody Seroconversion in Cancer Patients: Impact of Therapy Cessation-A Single-Center Study.

BACKGROUND: The effective development of COVID-19 vaccination has mitigated its harm. Using two laboratory methods, we investigated the efficacy of the BNT162b2 mRNA and BBIBP-CorV COVID-19 vaccines on seroconversion rates in cancer patients undergoing active cancer treatment. METHODS: SARS-CoV-2 vaccines were scheduled for 134 individuals. The consenting participants submitted three venous blood samples. Three samples: T0, T1, and T2. The ABBOTT-SARS-CoV-2 IgG II Quant and Elecsys® Anti-SARS-CoV-2 assays were used to evaluate the samples and convert the antibody titers to WHO (BAU)/mL units. RESULTS: Cancer patients exhibited a higher seroconversion rate at T2, regardless of vaccination type, and the mean antibody titers at T1 and T2 were higher than those at T0. BBIBP-CorV patients required a booster because BNT162b2 showed a higher seroconversion rate between T0 and T1. Statistics indicate that comparing Abbott and Roche quantitative antibody results without considering the sample collection time is inaccurate. CONCLUSIONS: COVID-19 vaccines can still induce a humoral immune response in patients undergoing cancer-targeted therapy. The strength of this study is the long-term monitoring of antibody levels after vaccination in cancer patients on active therapy using two different immunoassays. Further multicenter studies with a larger number of patients are required to validate these findings.

QuantiFERON-CMV and monitor predict cytomegalovirus, mortality, and graft-versus-host disease in transplant recipients.

Cytomegalovirus (CMV) is the most prevalent infection in recipients of hematopoietic stem cell transplant (HSCT). QuantiFERON-CMV (QF-CMV) and QuantiFERON-Monitor (QFM) assays were used to test whether immune-competent adult allogeneic HSCT recipients with CMV-specific T cells can control CMV infection or reactivation. Our data demonstrated a significant correlation between CMV infection measured by CMV-antigenemia test and QF-CMV results, graft versus host disease (GvHD), and mortality rates. The QF-CMV test revealed that CMV-specific T cells with higher interferon-γ (IFN-γ) release were correlated with lower CMV infection rates. There was a significant negative association between QF-CMV results, GvHD, and mortality rates. Data showed that a one-unit rise in IFN-γ was linked with a 12.7% reduction in GvHD and a 20.7% reduction in the mortality odds ratio. In addition, a negative correlation was found between QF-M results and CMV infection, with the QFM test predicting protection against CMV infection by 1.9%. This is one of the few studies establishing the QF-CMV test's predictive value for GvHD and mortality, its use to monitor HSCT patients for pre-emptive therapy, and the use of the QFM test to predict CMV infection and mortality in HSCT patients. Thus, these assays could be utilized to optimize preventive and pre-emptive therapy procedures to reduce transplant recipient adverse effects and posttransplant therapy costs.

Comparative Assessment of the Kinetics of Cellular and Humoral Immune Responses to COVID-19 Vaccination in Cancer Patients.

OBJECTIVE: The kinetics of immune responses to various SARS-CoV-2 vaccines in cancer patients were investigated. METHODS: In total, 57 cancer patients who received BNT162b2-RNA or BBIBP-CorV vaccines were enrolled. Cellular and humoral immunity were assessed at three-time points, before the first vaccine dose and 14-21 days after the first and second doses. Chemiluminescent microparticle immunoassay was used to evaluate SARS-CoV-2 anti-spike IgG response, and QuantiFERON® SARS-CoV-2 kit assessed T-cell response. RESULTS: Data showed that cancer patients' CD4+ and CD8+ T cell-median IFN-γ secretion of SARS-CoV-2 antigens increased after the first and second vaccine doses (p = 0.027 and p = 0.042). BNT162b2 vaccinees had significantly higher IFN-γ levels to CD4+ and CD8+ T cell epitopes than BBIBP-CorV vaccinees (p = 0.028). There was a positive correlation between IgG antibody titer and T cell response regardless of vaccine type (p < 0.05). CONCLUSIONS: This study is one of the first to investigate cellular and humoral immune responses to SARS-CoV-2 immunization in cancer patients on active therapy after each vaccine dose. COVID-19 immunizations helped cancer patients develop an effective immune response. Understanding the cellular and humoral immune response to COVID-19 in cancer patients undergoing active treatment is necessary to improve vaccines and avoid future SARS pandemics.

PD-L1 Expression in Colorectal Carcinoma: A Comparison of 3 Scoring Methods in a Cohort of Jordanian Patients.

Anti-programmed death-ligand 1 (PD-L1) treatments can improve colorectal carcinoma (CRC) survival; however, there is still controversy regarding the relationship between PD-L1 expression and the outcome of immunotherapeutic treatment and survival. The discrepancies are partly caused by the lack of a unified scoring system. This retrospective, cross-sectional study evaluated PD-L1 by immunohistochemistry in 127 CRC cases and compared the 3 scoring systems used to assess PD-L1: Tumor Percentage Score (TPS), Combined Positive Score (CPS), and immune cell (IC) score. Correlations were calculated using the χ 2 test. Kaplan-Meier curves with the Log-rank test were used to measure the contribution of PD-L1 expression to survival. PD-L1-positive rate were 29.9%, 57.5%, and 55.9% based on TPS, CPS, and IC score, respectively. TPS showed a better correlation with the clinicopathologic features being significantly higher with young age, T4, and adenocarcinomas (compared with mucinous/signet ring). TPS also showed an increasing trend with higher grade, lymph node stage, and male sex, although these variables were not significantly associated with PD-L1 expression. There was no correlation between PD-L1 expression and mismatch repair protein status in the 3 scoring methods. The probability of survival was higher for PD-L1-negative cases in the first 60 months after surgery if scored by the TPS method ( P =0.058). Future efforts correlating PD-L1 status with response to treatment are needed to decide on the best scoring method to be used for making therapy decisions.

Non-Dexamethasone Corticosteroid Therapy's Effect on COVID-19 Prognosis in Cancer Patients: A Retrospective Study.

BACKGROUND: Anti-inflammatory corticosteroids are used in cancer treatment and COVID-19 infections. Data on the impact of non-dexamethasone corticosteroids on COVID-19 infection severity in cancer patients are minimal. This study investigates if corticosteroid treatment affects the disease severity in adult cancer patients. METHODS: A total of 116 COVID-19-infected cancer patients on hydrocortisone (H) or prednisone (P) were compared to 343 untreated patients. The study included patients who received corticosteroids before (B), after (A), or both before and after (B and A) COVID-19 infections. Ventilation support, hospitalization and mortality were investigated. RESULTS: Our data showed that a significantly greater number of patients taking H or P required ventilation support and hospitalization and that mortality rates were higher than the control group. Patients who received H or P after COVID-19 infection had a significantly worse prognosis than the other sub-groups and the control group. CONCLUSION: Corticosteroids impacted cancer patients' COVID-19 prognosis. Despite the limited sample size, H- and P-treated patients' corticosteroids performed worse than the control, especially if treatments were received after COVID-19 infection. Hence, when a cancer patient already on H or P treatment is diagnosed with COVID-19, we recommend switching to a steroid treatment as suggested by international guidelines.

Programmed death-ligand 1 expression in diffuse large B-cell lymphoma is associated with poor prognosis.

BACKGROUND: Programmed death-ligand 1 (PD-L1) expression in some tumors has prognostic implications. This work aims at investigating PD-L1 expression in diffuse large B-cell lymphoma (DLBCL) and to study its association with clinicopathological variables. METHODS: The study consisted of 75 DLBCL patients who were cared for at the King Hussein Cancer Center during the period 2015-2018. The expression of PD-L1 in tumor tissue was assessed by immunohistochemistry using the anti-human PD-L1 (Clone 22C3) monoclonal antibody. The correlation between gender, age, clinical stage, pre-treatment-LDH level, tumor location, response to therapy, overall and event-free survival with PD-L1 expression was studied. RESULTS: Six patients were excluded from further analysis as they were in relapse at the time of tissue sampling. The tumor proportion score (TPS) was ≥1% in 16/69 (23.2%) of DLBCL cases while the combined positive score (CPS) at a cut-off of ≥20 was observed in 23/69 (33.3%) cases. No significant difference in PD-L1 expression was found between germinal center B-cell-like (GCB) and non-GCB subtypes. Similarly, no differences in PD-L1 expression (at CPS ≥20 and TPS ≥1) were found between different genders, age groups, clinical stages, tumor location, and patient response to therapy. However, base-line lactate dehydrogenase was significantly elevated in patients with PD-L1 CPS ≥20. The overall survival was not significantly different between PD-L1-positive and -negative groups. On the other hand, the median event-free survival was higher in either of the PD-L1 TPS or CPS negative groups at 107months each versus 54 months in the PD-L1 positive group of either category. CONCLUSIONS: PD-L1 expression can predict event-free survival in DLBCL cases and therefore poor prognosis.

A Rare Case of Nasal Sarcoma with BCOR Internal Tandem Duplication Showing Complete Pathologic Response to the VDC-IE Chemotherapy Protocol.

Sarcoma with BCOR genetic alteration is an exceptionally rare and emerging subtype of sarcoma. It is categorized into two types: BCOR-related gene fusions such as BCOR::CCNB3 sarcomas and other BCOR-rearranged sarcoma and sarcomas with internal tandem duplication of BCOR genes such as infantile undifferentiated round cell sarcomas and primitive myxoid mesenchymal tumors of infancy. BCOR::CCNB3 sarcomas predominantly arise in bone rather than soft tissue and exhibit a higher occurrence in children and adolescent males, whereas sarcomas with BCOR internal tandem duplication show a wider age range but usually arise in the first year of life. Due to their rarity, there is ongoing debate and uncertainty regarding the best treatment approach, with a lack of specific clinical trials addressing these tumors. In this report, we present a unique case of sarcoma with internal tandem duplication of BCOR gene originating in the nasal region. The tumor was successfully and completely resected using the standard VDC-IE chemotherapy protocol, resulting in an unprecedented 100 percent tumor necrosis. The patient has completed the protocol and remains recurrence-free 13 months after diagnosis. This case suggests potential efficacy of the standard VDC-IE protocol in achieving remarkable responses in BCOR rearrangement sarcomas, including the internal tandem duplication subtype. However, further studies are needed to determine the optimal treatment strategies for this disease.

The Effect of Dexamethasone Treatment on COVID-19 Prognosis in Cancer Patients.

Background: Dexamethasone is used to treat cancer, relieve chemotherapy-induced nausea and vomiting, enhance cancer patients' appetites, and treat COVID-19 patients. There is little evidence of the impact of a dexamethasone treatment plan on the severity of COVID-19 infections in cancer patients. This study explores whether dexamethasone treatment plan influences the severity of COVID-19 in dexamethasone-treated cancer patients. Methods: The medical records of 108 cancer patients receiving dexamethasone at King Hussein Cancer Center with a COVID-19 infection and 343 without corticosteroid treatment were reviewed. Patients on dexamethasone within seven days before infection, after infection, or both were included. Ventilation support, hospitalization, and mortality within 28 days of a COVID-19 diagnosis were key severity factors. Results: We found that dexamethasone before a COVID-19 infection increased the risk of requiring ventilation assistance and mortality within 28 days by a factor of 5.8 (2.8−12.0) relative to control (p < 0.005). Continuing dexamethasone treatment after a COVID-19 infection, or starting it after infection, had a risk factor equivalent to control. Conclusion: Our data showed that dexamethasone therapy protocol affected COVID-19 prognoses in cancer patients, and it is preferable to not discontinue therapy after infection. A rigorous prospective comparison between early and late dexamethasone dosing is needed to determine the best protocol for treatment.

An update on the impact of SARS-CoV-2 pandemic public awareness on cancer patients' COVID-19 vaccine compliance: Outcomes and recommendations.

Background: Aside from the pandemic's negative health effects, the world was confronted with public confusion since proper communication and favorable decisions became an ongoing challenge. As a result, the public's perceptions were influenced by what they knew, the many sources of COVID-19 information, and how they interpreted it. With cancer patients continuing to oppose COVID-19 vaccines, we sought to investigate the COVID-19 pandemic and vaccine sources of this information in adult cancer patients, which either helped or prevented them from taking the vaccine. We also assessed the relevance and impact of their oncologists' recommendations in encouraging them to take the vaccine. Methods: From June to October 2021, an online survey was conducted at King Hussein Cancer Center. A total of 441 adult cancer patients took part in the study. Patients who had granted their consent were requested to complete an online questionnaire, which was collected using the SurveyMonkey questionnaire online platform. Descriptive analysis was done for all variables. The association between categorical and continuous variables was assessed using the Pearson Chi-square and Fisher Exact. Results: Our results showed that 75% of the patients registered for the COVID-19 vaccine, while 12% refused vaccination. The majority of participants acquired their information from news and television shows, whereas (138/441) got their information through World Health Organization websites. Because the SARS-CoV-2 vaccines were made in such a short period, 54.7 % assumed the vaccines were unsafe. Only 49% of the patients said their oncologists had informed them about the benefits of SARS-CoV-2 vaccines. Conclusions: We found that SARS-CoV-2 vaccine hesitancy in cancer patients might be related to misinformation obtained from social media despite the availability of supportive scientific information on the vaccine's benefits from the physicians. To combat misleading and unreliable social media news, we recommend that physicians use telehealth technology to reach out to their patients in addition to their face-to-face consultation, which delivers comprehensive, clear, and high-quality digital services that guide and help patients to better understand the advantages of COVID-19 vaccines.

Establishing the First COVID-19 Convalescent Plasma Biobank in Jordan.

Background: Antibodies with the specialized ability to fight infection can be found in the blood of individuals who have recovered from or have been vaccinated against COVID-19. As a result, plasma from these individuals could be used to treat critically ill patients. This treatment is known as convalescent plasma (CCP) therapy. Methods: Plasma units from 1555 consented healthy blood bank donors were collected from February to September 2021. Blood units were tested for the quantitative determination of Immunoglobulin G (IgG) antibodies to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus using one of the following assays based on the availability of the kits: The LIAISON® SARS-CoV-2 TrimericS IgG assay or the Abbott SARS-CoV-2 IgG II Quant assay. Results: Among the tested donors, 1027 participants tested positive for neutralizing anti-SARS-CoV-2 IgG antibodies (66.04%). There were 484 donors whose plasma qualified to be used for CCP therapy (47.13%) and 214 CCP units were stored in the COVID-19 convalescent biobank. Conclusion: We were able to identify and store 214 fresh frozen plasma units qualified for CCP-plasma therapy for COVID-19 patients according to World Health Organization standards. Hence, we established the first COVID-19-convalescent plasma data and plasma biobank for treating COVID-19-infected cancer patients in Jordan and the region.

Anaplastic lymphoma kinase rearrangements in patients with non-small cell lung cancer in Jordan.

OBJECTIVE: Anaplastic lymphoma kinase (ALK) rearrangement is an important oncogenic driver in some non-small cell lung cancers (NSCLC). Treatment with ALK tyrosine kinase inhibitors improves survival. The availability of diagnostic immunohistochemistry (IHC) has led to a paradigm shift in ALK testing. This study examined the prevalence of ALK rearrangement in Jordanian patients with NSCLC and compared the results of IHC and fluorescence in situ hybridization (FISH) for detecting ALK rearrangement. METHODS: This retrospective study on 449 patients with NSCLC treated at the King Hussein Cancer Center in Jordan tested biopsy samples for ALK rearrangement using FISH and/or IHC (D5F3) between 2018 and 2020. RESULTS: Eighteen patients (4%) had ALK-positive NSCLC. The calculated sensitivity and specificity of ALK immunostaining compared with FISH were 87.5% and 96%, respectively. ALK-positive patients were significantly younger than their ALK-negative counterparts, and women were three times more likely to carry ALK rearrangement than men. ALK rearrangement was significantly associated with smoking history, with most ALK-positive patients being non-smokers, former smokers, or light smokers. CONCLUSIONS: IHC is a reasonable alternative to FISH for ALK testing with advantages in terms of robustness, turnaround times, and cost-effectiveness.

Comparison of the Immunogenicity and Protective Efficacy of Various SARS-CoV-2 Vaccines among Healthcare Workers: Are Our White Coat Armies Protected?

Background: The effective immunization of healthcare workers (HCWs) plays a vital role in preventing the spread of SARS-CoV-2 infection during the coronavirus disease 2019 (COVID-19) pandemic. There is limited data on the immune response to vaccination among HCWs. We aim to determine seroprevalence rates and neutralizing IgG antibody response to various immunizations among HCWs. Methods: This study was conducted between July and September 2021, in which blood samples were obtained from HCWs and SARS-CoV-2 IgG neutralizing antibodies were measured. Data regarding vaccination status with Pfizer/BioNTech, Sinopharm, or AstraZeneca vaccines, occupation, and prior COVID-19 infection were analyzed. Results: COVID-19 infection post-vaccination was associated with higher mean antibody titers, regardless of vaccine type. Pfizer/BioNTech vaccination produced higher mean antibody titers for HCWs with prior COVID-19 infection (p < 0.00001) than other types of vaccines. Although 96% of HCWs were vaccinated, 3% were seronegative. For HCWs who were seropositive, there were no significant differences between the mean antibody titers when comparing occupations and blood indices. Conclusion: Awareness of the immunity status of HCWs is key to protecting this important group against SARS-CoV-2, especially those without prior COVID-19 infection. Further public health efforts regarding booster vaccination for HCWs are crucial to provide necessary antibody protection.

The clinicopathological features of programmed death ligand-1 expression in colorectal carcinoma.

BACKGROUND: Few studies have addressed the clinicopathological features of colorectal cancer (CRC) that express programed death-ligand 1 (PD-L1). Various assays and scoring methodologies were used and thus inconsistent results were obtained. In this study, we aimed to investigate the relationship of PD-L1 expression in CRC with various clinicopathological variables using a standardized assay and scoring algorithm. DESIGN: Tissue microarrays were constructed from 91 random cases of CRC diagnosed at King Hussein Cancer Center (KHCC). Immunohistochemical (IHC) staining using the monoclonal antibody 22C3 was performed. Scoring using the standard "Combined Positive Score" (CPS) method was done. CPS of ≥1 was considered positive. Various clinicopathological features were collected from the medical records of the patients. RESULTS: Of the 91 cases, 49 (53.8%) were PD-L1 positive (CPS ≥1). PD-L1 expression was more frequent among moderately differentiated carcinomas (43 of 72 (59.7%) were positive compared to 6 of 19 (31.5%) poorly differentiated cases (P = 0.029)); among node negative cases (21 of 24 (87.5%) N0 cases were PD-L1 positive in contrast to 28 of 67 (41.8%) N1/N2 cases (P = <0.001)); among mucinous subtype (12 of 15 (80%) of cases (P = 0.02)); and among mismatch repair deficient (dMMR) (16 of 16 (100%) versus 11 of 30 (36.6%) MMR proficient (P = <0.001)). Age, gender, localization, and T or M stages were not significantly associated with PD-L1 expression. CONCLUSION: PD-L1 expression in CRC is associated with favorable prognostic features; namely, lower grade, N0, mucinous variant, and dMMR tumors.

Changing Trends in Estrogen Receptors/Progesterone Receptors/Human Epidermal Growth Factor Receptor 2 Prevalence Rates Among Jordanian Patients With Breast Cancer Over the Years.

PURPOSE: Estrogen receptors (ERs), progesterone receptors (PRs), and human epidermal growth factor receptor 2 (HER2) are the mainstay of breast cancer management, and their prevalence rates vary among different populations possibly related to ethnic/genetic and/or socioeconomic status. In a previous study conducted at the King Hussein Cancer Center (published 2006), Jordan ER/PR/HER2 rates for patients diagnosed in 2003-2004 were 50.8%/57.5%/17.5%, respectively. The aim of this study is to revisit the prevalence rates to see if they have changed over the years with changing socioeconomic status. MATERIALS AND METHODS: We retrieved clinicopathologic data of all patients (1,185) diagnosed with breast cancer during 2018. The data included age, histologic type, grade, and ER/PR/HER2 status as determined by immunohistochemistry and/or fluorescence in situ hybridization for HER2. RESULTS: The mean age of patients was 52 (median = 51, range = 25-92) years, and the majority (73.2%) had invasive carcinoma of no special type. ER/PR/HER2 were 77.0%/72.4%./23.8%, respectively. Triple-negative breast cancers were 10.1%. In comparison with previous results of 2006, the changes are statistically significant. Similar changes were seen in other Middle Eastern populations. The current rates are close to those of Western populations. CONCLUSION: Rates of ER/PR/HER2 expression have significantly changed and are close to those of Western populations for ER/PR. We propose that such changes are secondary to the adoption of a westernized lifestyle and socioeconomic changes.

Changing trends in seroprevalence rates of transfusion-transmitted diseases among blood donors in Jordan.

BACKGROUND: Hepatitis B and C infections and transmission are a serious challenge to all healthcare systems. We studied seroprevalence rates of Transfusion Transmitted Diseases (TTD) among blood bank donors in Jordan from 2014 to 2019 as a follow-up study of our previously published work. In addition, we wanted to explore the efficacy of the mandatory vaccination of infants against hepatitis B virus (HBV) which was implemented by the Ministry of Health since 1995 for the eradication of HBV infection in Jordan. METHODS: We reviewed blood bank donors' records at King Hussein Cancer Center (KHCC) from January 1st, 2014, until December 31st, 2019. Results of seropositivity prevalence rates for HBsAg, anti-HBcore, and anti-HCV, using Enzyme-Linked ImmunoSorbent Assay (ELISA) were compared to seropositivity rates from our previously published data. In addition, our results were compared to data obtained from other blood banks in Jordan, as well as compared to published information from blood banks in neighboring countries. RESULTS: The prevalence rates (%) of seropositive blood donors for viral hepatitis for the years 2014, 2015, 2016, 2017, 2018, and 2019, were as follows: HBsAg rates were 0.3386, 0.2108, 0.1801, 0.1898, 0.2068, and 0.2741; anti-HBcore rates were 4.1112, 3.2271, 2.9748, 2.8405, 2.6879 and 3.0986; and anti-HCV rates were 0.1129, 0.0486, 0.0548, 0.0654, 0.0782, and 0.0839, respectively. There was a significant increase in the prevalence of HBsAg, Anti-HBcore and Anti-HCV antibodies in 2019 (one sample z-score test, p < 0.00001). CONCLUSIONS: Prevalence rates of hepatitis B and C infections among Jordanian blood bank donors showed a steady decline between 2009 and 2017, and these rates were much lower in Jordan than in neighboring countries. However, an increase in the prevalence rates of hepatitis B and C infections among blood bank donors was documented in 2019. While the reasons for this increase are not clear yet, these findings highlight the importance of renewed efforts to increase public health awareness of HBV and implement effective measures to prevent the transmission and infection with HBV, including national vaccination programs.

Dramatic rise in seroprevalence rates of SARS-CoV-2 antibodies among healthy blood donors: The evolution of a pandemic.

BACKGROUND: Seroprevalence studies of SARS-CoV-2 antibodies are useful in assessing the epidemiological status in the community, and the degree of spread. OBJECTIVE: To study the seroprevalence rates of SARS-CoV-2 antibodies among healthy blood donors in Jordan, at various points of time and as the pandemic evolves in the community. METHODS: In total, 1374 blood donor samples, from three groups, were tested for SARS-CoV-2 total immunoglobulin antibodies. In the first group, samples from 734 individuals (from donations made between January and June 2020) were tested in June. In the second group, 348 individuals were tested in September 2020. The third group of 292 individuals was tested in February 2021. A qualitative assay was used for testing (specificity 99.8%, sensitivity 100%). RESULTS: The first two groups, from January-June and September 2020, when confirmed Covid-19 cases numbered between several hundred and 3000, showed a seroprevalence rate of 0% (95% CI 0.00-0.51%). The third group (early February 2021), when the number of confirmed cases had reached 100 times that of September 2020, revealed a seroprevalence of 27.4% (95% CI 22.5-32.9%). CONCLUSIONS: A dramatic rise in seroprevalence of SARS-CoV-2 antibodies was seen among healthy blood donors in Jordan, in parallel with widespread intracommunity transmission of the disease. This information is useful for assessing the degree of herd immunity, and provides for better understanding of the pandemic.

PD-L1 Expression Harmonization in Gastric Cancer Using 22C3 PharmDx and SP263 Assays.

The immune checkpoint inhibitor Pembrolizumab has been FDA-approved for the treatment of gastric cancer (GC) and gastroesophageal junction (GEJ) cancer in patients who fail second-line therapy and test positive by a companion programed death ligand 1 (PD-L1) assay, the 22C3 PharmDx. It would be useful to investigate the potential interchangeability of other PD-L1 assays in order to develop a more sustainable diagnostic strategy. We investigated the possibility of harmonizing different PD-L1 assays, utilizing samples from 94 GC and GEJ patients to compare their expression using 2 laboratory developed tests (LDTs): The Dako 22C3 antibody and the Ventana SP263 run on the Ventana platform with the FDA-approved companion diagnostic test, the 22C3 PharmDx. This would be the first report assessing the 22C3 on Ventana's platform in GC. Pearson correlation coefficients between the Dako 22C3 PharmDx and the 22C3-LDT and the Ventana SP263 assays were 0.965 (P<0.001) and 0.932 (P<0.001), respectively, which indicates an almost perfect correlation. The sensitivity and specificity were also high at different cutoffs [both 100% at combined positive score (CPS)≥1 and 92.59% and 95.52% at CPS≥10, respectively] for the comparison between Dako 22C3/22C3-LDT assays. As for the sensitivity and specificity between the Dako 22C3/Ventana SP263 assays the results were 100% and 95.67% at CPS≥1; and 96.30% and 95.52% at CPS≥10, respectively. In conclusion, the analytical performance of 22C3 and SP263 clones on the Ventana platform was close to that of the reference assay (Dako 22C3 assay), suggesting that the 2 LDTs can be utilized interchangeably with the FDA-approved standard assay as an aid to select GC and GEJ patients for Pembrolizumab treatment.

Expression of PD-L1 using SP142 CDx in triple negative breast cancer.

Triple negative breast cancer (TNBC) represents a small subtype of breast cancer yet it has the worst outcome. Immunotherapy using immune checkpoint inhibitors combined with chemotherapy was recently approved by the FDA raising the hope for an improved outcome. The approval was based on demonstration of a positive PD-L1 expression using the SP142 CDx assay 1. We aimed to study a cohort of TNBC patients in terms of prevalence of the PD-L1 expression using the approved assay and to investigate its association with clinicopathological variables. This is a single center retrospective study consisting of 49 TNBC patients who had available archived paraffin-embedded tissue blocks from the primary tumors. All blocks were stained using the SP142 CDx assay as per the manufacture's instruction. Clinicopathological data were collected from medical records. Eighteen of the 49 (36.7%) patients were found to have a score of 1% or more by the immune cell-scoring algorithm. PDL-1 expression was significantly associated with the degree of tumor infiltrating lymphocytes (TILs). No additional significant relationship was found between PD-L1 expression and any of the other investigated clinicopathological variables. Although a trend of favorable prognostic association with PD-L1 expression was noted. The overall and event free survival were significantly related to pathological response to neoadjuvant therapy. Conclusion: Our PD-L1 rate of 36.7% is close to the results of the previously reported 40.9% in the IMpassion 130 trial. There were no significant association between positive PD-L1 expression and clinicopathological variables however a trend of a favorable outcome was observed.

Comparison of Estrogen receptors, Progesterone receptors and HER2-neu immunohistochemistry results in breast cancer with those of Oncotype Dx.

Oncotype Dx (ODx) recurrence score (RS) is used in early breast cancer to guide the use of adjuvant therapy. In addition to RS the test produces results of reverse transcriptase-polymerase chain reaction (RT-PCR) of Estrogen receptors (ER), Progesterone receptors (PgR) and Human epidermal growth factor receptor 2 (HER2-neu). Our goal was to determine the correlation between immunohistochemistry (IHC) and RT-PCR testing of ER, PgR and HER2-neu and to correlate the results of ODx RS with tumors' grade, age and PgR status. 113 patients with ER+, HER2-neu- breast cancers that underwent ODx testing were analyzed for receptors correlation and concordance rates by the 2 methods. A total of 104 patients had ER+/PgR+ tumors and 9 patients had ER+/PgR- tumors by IHC, the average RS were 17.5 ± 9.1 and 31.2 ± 8.7 (P < 0.001) respectively. The Spearman correlation coefficient between IHC and ODx results were 0.5 (95% CI 0.34-0.62) for ER and 0.78 (95% CI 0.7-0.84) for PgR. The concordance rate between IHC and ODx was 98.2% for ER, 89.4%.for PgR and 99.1% for HER2-neu. Most of the discordant cases (9 out of 13) were low positive (1-10%) by IHC and negative by RT-PCR. In addition higher tumor grade was associated with a higher ODx RS. Our data show that the IHC results were highly concordant with RT-PCR for ER, PgR and Her2-neu. In addition low positive (1-10%) ER/PgR might indicate a real negative status. Our study shows that ER+/PgR- breast cancers are associated with a significantly higher ODx RS.