CD Markers Research Articles

Investigating T Cell Immune Dynamics and IL-6's Duality in a Microfluidic Lung Tumor Model.

Interleukin 6 (IL-6), produced by immune cells, is crucial in promoting T cell trafficking to infection and inflammation sites, influencing various physiological and pathological processes. Concentrations of IL-6 and other cytokines and chemokines can influence T cell differentiation and activation. Understanding the dual faces of IL-6 within the tumor microenvironment is crucial to understanding its role. A flow-based microsystem was designed to investigate CD4+ T cell activation in response to different IL-6 gradients in an under-control 3D culture. The study found that cancer cells' response to varying IL-6 concentrations was dynamic and dose-sensitive, with immune cell migration rates showing sensitivity to the IL-6 gradient. A549 cell expansion increases gradually and time-dependently with 50 ng of IL-6, while Jurkat cell migration follows a time-dependent pattern. However, when a total of 100 ng IL-6 concentration is applied, A549 cells expand rapidly, potentially influencing Jurkat cell migration. Jurkat cell mobility is lower, possibly due to increased A549 cell presence and heightened cell-cell interactions. Different IL-6 concentration gradients can modulate the expression of some CD markers like CD69 and programed cell death protein 1 in CD4+ T cells, suggesting that IL-6 concentration gradients affect immune cell phenotypes. This suggests that IL-6 plays a crucial role in activating T helper cells and may be involved in the later phases of inflammation. Also, the increased levels of IFN-γ and TNF-α highlight IL-6's impact on T cell inflammatory response. This study emphasizes the intricate effects of IL-6 on T cell activation, phenotype, cytokine production, and phenotypic heterogeneity, providing valuable insights into immune response modulation in an experimental setting.

Impact of Dolutegravir plus Lamivudine as First-Line Antiretroviral Treatment on HIV-1 Reservoir and Inflammatory Markers in Peripheral Blood.

To compare the effects of first-line antiretroviral treatment (ART) with dolutegravir plus lamivudine (DTG+3TC) versus DTG plus emtricitabine/tenofovir alafenamide (FTC/TAF) on the evolution of the HIV-1 reservoir and immune activation biomarkers in people with HIV (PWH). DUALITY was a 48-week, single-center, randomized, open-label clinical trial in ART-naïve PWH. Participants were randomized (1:1) to receive ART with DTG+3TC (2DR group) or DTG+FTC/TAF (3DR group). Total and intact proviral HIV-1 DNA, cell-associated RNA in CD4+ T cells, the frequency of HIV-infected CD4+ T cells able to produce p24, plasma soluble inflammatory markers (IL-6, sCD14, TRAIL, IP-10, FABP2, CRP and D-dimer), and activation and exhaustion markers in CD4+ and CD8+ T cells were longitudinally determined. Forty-four participants (22 per study arm) were enrolled. Baseline mean (SD) log10 plasma viral load (pVL) and CD4+ T cell counts were 4.4 (0.7) copies/mL and 493 (221) cells/mm3, respectively. All participants completing the study (2DR n=20; 3DR n=21) had pVL <50 copies/mL at week 48, except for one in the 2DR group who was resuppressed after treating syphilis. Changes from baseline to week 48 in all reservoir parameters or in levels of soluble inflammatory biomarkers and activated or exhausted CD4+ and CD8+ T cells were similar between 2DR and 3DR groups. First-line ART with DTG+3TC resulted in a similar reduction of HIV-1 persistence parameters in peripheral blood, and comparable changes in immune-associated soluble and T-cell markers compared with DTG+FTC/TAF. These findings support recommendation of DTG/3TC among preferred options for first-line ART in PWH.

Diagnosis of canine B-cell chronic lymphoid leukemia with a CD21 negative phenotype using the LT21 clone CD21 antibody in flow cytometry: a case report

BackgroundChronic lymphoid leukemia (CLL) is a hematological disorder characterized by the clonal expansion of small mature lymphocytes that accumulate in the blood and bone marrow. CLL can arise from B-, T-, or natural killer cell clones. The cytological evaluation of blood smears is often the simplest and least invasive method for diagnosing lymphoid leukemia. Immunophenotyping is used to further subclassify the type of lymphoid leukemia.Case presentationA 15-year-old, 4.4-kg spayed female Shih Tzu was presented to the veterinary medical teaching hospital of Kangwon National University. Despite having a normal appetite and activity level, cervical and inguinal lymph node enlargement was noted on physical examination. Complete blood count revealed severe leukocytosis, severe lymphocytosis, and monocytosis. Splenomegaly, hepatomegaly, and lymph node enlargement were detected on radiographic and ultrasonographic examination. Immunophenotyping was performed using peripheral blood mononuclear cells (PBMCs). The majority of lymphocytes exhibited the following profiles: CD3−CD79a− (97.5%), CD4−CD8− (98.6%), CD21−CD79a− (98.4%), CD34− (0.1%), CD45+ (99.6%), major histocompatibility complex class II+ (99.5%), and CD14− (0.5%). Based on the immunophenotyping results, possible differentials considered included the following: the majority of lymphocytes may be natural killer (NK) cell clones, plasma cell clones, or show aberrant expression or loss of CD21 marker due to the neoplastic nature of the cells. Further flow cytometry was performed using antibodies against CD3, CD5, CD94, and granzyme B. The combined results indicated that the predominant lymphocyte subset in the PBMCs was CD3−CD5−CD21−CD94−granzyme B−. To confirm monoclonality and exclude the aberrant loss of CD markers, a polymerase chain reaction for antigen receptor rearrangement (PARR) assay was conducted. The PARR assay, using DNA from blood and lymph node samples, showed B-cell monoclonality. Immunocytochemistry using PBMCs showed that the plasma cell marker Multiple Myeloma Oncogene 1 (MUM1) was not expressed. Therefore, the diagnosis was confirmed to be B-cell CLL.ConclusionImmunophenotyping can help subclassify the type of lymphoid leukemia; however, as tumor cells can show aberrant expression or loss of the CD21 marker, combining immunophenotyping with the PARR assay could yield a more accurate diagnosis.

The impact of prebiotic, probiotic, and synbiotic supplements on CD4, CD8 counts and inflammatory markers in HIV patients: A systematic review and meta-analysis

The impact of prebiotic, probiotic, and synbiotic supplements on CD4, CD8 counts and inflammatory markers in HIV patients: A systematic review and meta-analysis

Fractal Dimension Analysis of the Tumor Microenvironment in Cutaneous Squamous Cell Carcinoma: Insights into Angiogenesis and Immune Cell Infiltration

The global incidence of cutaneous squamous cell carcinoma (cSCC), a prevalent and aggressive skin cancer, has risen significantly, posing a substantial public health challenge. This study investigates the tumor microenvironment (TME) of cSCC by focusing on the spatial distribution patterns of immune and vascular markers (CD31, CD20, CD4, and CD8) using fractal dimension (FD) analysis. Our analysis encompassed 141 cases, including 100 invasive cSCCs and 41 specimens with pre-invasive lesions exclusively, and the rest were peripheral pre-invasive lesions from the invasive cSCC class. The FD values for each marker were computed and compared between pre-invasive and invasive lesion classes. The results revealed significant differences in FD values between the two classes for CD20 and CD31 markers, suggesting distinct alterations in B cell distribution and angiogenic activity during cSCC progression. However, CD4 and CD8 markers did not exhibit significant changes individually. Still, the CD4/CD8 ratio showed a significant difference, suggesting a potential shift in the balance between T helper and cytotoxic T cell responses, impacting the immune landscape as lesions progressed from pre-invasive to invasive stages. These findings underscore the complexity and heterogeneity of the TME in cSCC and highlight the potential of FD analysis as a quantitative tool for characterizing tumor progression. Further research is needed to elucidate the implications of these differences in the clinical management of cSCC.

Flow Cytometry Analysis of CD4⁺ and CD8⁺ T Lymphocytes in COVID- 19 Patients

The clinical presentation of COVID-19 varies greatly from case to another, with host characteristics playing a pivotal role. The immune system is a pivotal function in establishing the course of SARS-COV-2 infection. Severe instances of COVID-19 and the resulting death are caused less by the virus itself than by the destructive unrestrained immune response. Among the host determinants for poor disease prognosis is probably genetic vulnerability to dysregulated immune response. The objective of this work is to examine some of the host genetic variables that are being studied in relation to the severity of COVID-19. The complex relationship between autoimmunity and the pathophysiology of COVID-19 was examined. Patients with COVID-19 were recruited from AL-Dewaniyah Teaching Hospital and AL-Hussain Teaching Hospital in Karbala city. All patients had been diagnosed with the virus using real-time polymerase chain reaction. A total of 70 blood samples (35 from the mild group, 35 from the severe group, and 35 from the controlled group) were obtained in EDTA tubes. Orderly to investigate the correlation between the progression of disease and variations in specific regions of host immunoassays had been used by flow cytometry. The results of the flow cytometry were reported as the average percentage of positive cells for CD4 and CD8 immunological markers. Cases with COVID-19 had a substantially lower mean CD4 concentration than the healthy patients. Furthermore, the mean concentration of CD8 was significantly lower in cases with COVID-19 than in healthy. In addition, there is significant (P 0.001) decline in both CD4 and CD8 mean levels between patients with severe and mild COVID-19.

The frequency of CD38+ alveolar macrophages correlates with early control of M. tuberculosis in the murine lung

Tuberculosis, caused by Mycobacterium tuberculosis, remains an enduring global health challenge due to the limited efficacy of existing treatments. Although much research has focused on immune failure, the role of host macrophage biology in controlling the disease remains underappreciated. Here we show, through multi-modal single-cell RNA sequencing in a murine model, that different alveolar macrophage subsets play distinct roles in either advancing or controlling the disease. Initially, alveolar macrophages that are negative for the CD38 marker are the main infected population. As the infection progresses, CD38+ monocyte-derived and tissue-resident alveolar macrophages emerge as significant controllers of bacterial growth. These macrophages display a unique chromatin organization pre-infection, indicative of epigenetic priming for pro-inflammatory responses. Moreover, intranasal BCG immunization increases the numbers of CD38+ macrophages, enhancing their capability to restrict Mycobacterium tuberculosis growth. Our findings highlight the dynamic roles of alveolar macrophages in tuberculosis and open pathways for improved vaccines and therapies.

Immunoproteomics reveal different characteristics for the prognostic markers of intratumoral-infiltrating CD3+ T lymphocytes and immunoscore in colorectal cancer

Tumor-infiltrating lymphocytes (TILs) and immunoscoring based on densities of CD3+ and CD8+ TILs are both favorable prognostic markers in colorectal cancer (CRC). However, determination of the molecular features of TILs, particularly their immunoproteomic signatures would require the development of large-scale in situ spatiotemporal technologies. Recently, a multiplex in situ digital spatial proteomic profiling (DSP) tool GeoMx DSP has been applied to identify biomarkers predictive of therapeutic responses and to understand disease mechanisms and progression. Taking advantage of this tool, we simultaneously characterized the spatial distribution and interactions of 42 immune proteins in tumor cells (TC), CD3+ T stromal TILs (sTILs), and CD20+ B sTILs using tissue microarrays, and further studied their associations with CD3+ T TILs and immunoscores in CRC. First, our data showed that well-known immune checkpoints, such as PD-L1, PD-L2, and LAG3, were expressed at low levels, whereas some other immune proteins, such as CD11c, CD68, STING, and CD44, were highly expressed. Second, eight spatial interactions were identified, including five interactions between TC and CD20+ B sTILs, two interactions between CD3+ T sTILs and CD20+ B sTILs, and one interaction among TC, CD3+ T sTILs, and CD20+ B sTILs. Third, the differential immune microlandscape in the spatial compartments was identified in tissues with positive CD3+ T intratumoral TILs and high immunoscores. Collectively, our study is the first to provide in situ spatial immune characteristics at the proteomic level. Moreover, our findings provide direct evidence supporting the infiltration of CD3+ T sTILs from stoma to TC and shed important insights into better understanding and treating CRC patients related to different immune prognostic markers.

Bojungikki-Tang Augments Pembrolizumab Efficacy in Human PBMC-Injected H460 Tumor-Bearing Mice

Bojungikki-Tang (BJIKT) is traditionally used to enhance digestive function and immunity. It has gained attention as a supplement to chemotherapy or targeted therapy owing to its immune-boosting properties. This study aimed to evaluate the synergistic anti-tumor effects of BJIKT in combination with pembrolizumab in a preclinical model. MHC I/II double knockout NSG mice were humanized with peripheral blood mononuclear cells (PBMCs) and injected subcutaneously with H460 lung tumor cells to establish a humanized tumor model. Both agents were administered to evaluate their impact on tumor growth and immune cell behavior. Immunohistochemistry showed decreased exhaustion markers in CD8(+) and CD4(+) T cells within the tumor, indicating enhanced T cell activity. Additionally, RNA sequencing, transcriptome analysis, and quantitative PCR analysis were performed on tumor tissues to investigate the molecular mechanisms underlying the observed effects. The results confirmed that BJIKT improved T cell function and tumor necrosis factor signaling while suppressing transforming growth factor-β signaling. This modulation led to cell cycle arrest and apoptosis. These findings demonstrate that BJIKT, when combined with pembrolizumab, produces significant anti-tumor effects by altering immune pathways and enhancing the anti-tumor immune response. This study provides valuable insights into the role of BJIKT in the tumor microenvironment and its potential to improve therapeutic outcomes.

Immune cells and tryptophan metabolism in the joint capsule tissue in rheumatoid arthritis

To the present day, many links in the pathogenesis of rheumatoid arthritis remain unclear, which leads to unsatisfactory results in its therapy.The aim. To study the cells involved in immune reactions and tryptophan metabolites in the joint capsule in rheumatoid arthritis.Materials and methods. The experiments were carried out on 40 Wistar rats. Rheumatoid arthritis was induced by intraperitoneal injection of a solution of type 2 collagen (Chondrex Inc., USA) in incomplete Freund’s adjuvant. On the days 7, 14 and 21, the content of tryptophan, kynurenine, 3-hydrokenurinine, L-5-hydrotryptophan in the joint capsule was determined using high-performance liquid chromatography. Cells with CD3, CD20 and CD68 in joint tissues were studied at the same time using the streptavidin-biotin-peroxidase method. We used enzyme-linked immunosorbent method to determine antibodies to citrulline-containing peptide. Statistical analysis was performed using the Jamovi, version 2.3 software.Results. The content of cells carrying CD3, CD20 and CD68 markers in the joint was high in experimental rheumatoid arthritis. In joint tissues, the content of tryptophan metabolites along the kynurenine pathway also increases and the concentration of metabolites along the serotonin pathway decreases. Direct positive correlations of cells carrying CD3, CD20 and CD68 differential clusters with the content of tryptophan metabolites along the kynurenine pathway and negative correlations with metabolites of the serotonin pathway were established.Conclusions. Cells carrying CD3, CD20 and CD68 markers and tryptophan metabolites – kynurenine and L-5-hydrotryptophan – play an important role in the pathogenesis of rheumatoid arthritis.

Effect of tetrapeptides KK1 and KK5 on the immunological parameters of rat spleen during passive smoking

One of the trends in experimental and clinical immunology is the creation of new immunomodulators aimed at correcting various variants of the pathology of the immune system. Of interest are non-toxic, hormone-free and protease-resistant tetrapeptides, homologue of the of adrenocorticotropic hormone fragment (15- 18). There is little data in the literature on the effect of the above immunomodulators on the immunological parameters of the rat spleen on xenobiotic exposure models, which makes research relevant in this aspect. For the first time, the effect of tetrapeptides with laboratory ciphers KK1 and KK5 on the immunological parameters of the spleen of 36 female Wistar rats with passive smoking was evaluated. The experimental rats were fumigated with tobacco smoke for 8 hours daily for 20 days. Synthetic tetrapeptides were administered intranasally at a dose of 40 micrograms per kg/day, starting from the 10th day of the experiment. It was found that the studied tetrapeptides had a unidirectional positive effect on the immunological parameters of the spleen of experimental animals with passive tobacco smoking. This was expressed in a tendency to restore the pool of cells with CD3, CD4, CD8, CD20 markers to the level of the control group, as well as spontaneous and induced cytokine production by splenocytes. The shifts in the parameters of the spleen identified in this work may be based on a number of possible causes. Firstly, passive smoke causes a stress response in rats, which is confirmed by previous studies on a similar model, on an increase in the level of stress hormones in the blood serum of Wistar rats. Secondly, the literature data indicate the activation of lipid peroxidation during smoking. Thirdly, it is known that under the action of tobacco smoke toxicants, the lymphoid cell line suffers the most, since their polyhydrooxidized metabolites accumulate in the bone marrow and lymphoid organs, causing hypoplasia of the central and peripheral organs of immunity. A visible sign of this phenomenon is a decrease in the cellularity in the spleen, which is established in this work. Considering that active and passive smoking causes hypoxia and increases lipid peroxidation, the use of tetrapeptides KK1 and KK5 as agents that improve the body’s adaptation to hypoxia and increase resistance to stress damage is justified. Thus, the tetrapeptides KK1 and KK5 have a positive immunomodulatory effect, reducing the toxic stress effects of passive smoking.

Expression of immune checkpoint markers in peripheral blood lymphocytes of women with fibroadenoma of the breast

Changes of the immune state in patients with fibroadenoma (FA) or other benign lesions of the breast, as well as the involvement of immune checkpoints in pathogenesis of these lesions, remain underexplored. The aim of this study was to compare the expression level of the key immune checkpoint markers CD279/PD-1, CD274/PD-L1, CD366/TIM3, and CD223/LAG3 in total circulating lymphocytes, T cell population as well as in its CD4 and CD8 subsets in peripheral blood from women with breast FA and healthy controls. Blood samples were taken from 12 women diagnosed with FA of the breast (aged 23-54 years, FA group) and 15 healthy women (aged 22-52 years, control group). Sample uptake was performed immediately before surgery, and samples were further analyzed by multicolor flow cytometry using monoclonal antibodies CD3-VioBlue, CD4/CD8-FITC, PD1-PE, PD-L1-PerCP-Cy.5.5, and TIM3/LAG3-APC. Each sample was incubated with 4 antibody combinations: CD3/CD4/PD1/PD-L1/TIM3, CD3/CD4/PD1/PD-L1/LAG3, CD3/CD8/PD1/PD-L1/TIM3, and CD3/CD8/PD1/PD-L1/LAG3. First, mono-expression of each of the 4 immune checkpoint markers was evaluated in the lymphocyte gate from both investigated groups. In FA samples, a significant increase in PD-L1 expression (assessed as percent of positive cells and fluorescence intensity change) was observed. Regarding expression of immune checkpoints in CD3+ T cells, along with significantly increased %PD-L1+, elevated numbers of PD1+T cells were detected. As for the differences in immune checkpoint expression changes between CD4+ and CD8+ T cell subsets, FA patient group demonstrated a more prominent increase in the amount of CD8+PD1+T cells relative to CD4 subset. The profiles of PD-L1 changes in CD4 and CD8 subpopulations were comparable showing, in both cases, a significant increase in FA sample group. We also analyzed changes in co-expression of any two immune checkpoint markers in CD4+ and CD8+ T cell subsets. The most noticeable was as increase in the prevalence of PD1+PD-L1+ phenotype in both T cell subpopulations from FA patients compared to the controls. With respect to co-expression of 3 checkpoint markers in CD4+ and CD8+ T cells, a significant increase in the percentage of PD1+PD-L1+TIM3+ cells among CD4 T helpers was found in FA. Thus, specific changes of T cell phenotype related to (co-)expression of immune checkpoint regulators may occur in systemic circulation of women with breast FA.

Immunological Tumor Microenvironment of Solitary Fibrous Tumors-Associating Immune Infiltrate with Variables of Prognostic Significance.

Solitary fibrous tumors (SFTs) are morphologically heterogeneous tumors characterized by the NAB2::STAT6 gene fusion. Clinical outcomes may vary widely, and while most cases have favorable outcomes, some can progress to aggressive disease, manifesting as recurrence and metastasis, and ultimately resulting in patient death. Herein, we analyze the immunological tumor microenvironment (ITME) of SFTs, aiming to determine its prognostic value and correlation with established risk stratification systems (RSSs). A retrospective observational multicenter study of 52 fusion-confirmed SFTs with clinical follow-up data. Immunohistochemical analysis including CD163, CD68, CD3, CD8, CD20, PDL-1, PD-1, and LAG1 were evaluated in tissue microarrays, using an analog scale with scores ranging from 0 to 3 (0 = ≤9, 1 = 10-49, 2 = 50-99, and 3 = >100 positive cells per 10 high-power fields). The expression of these markers was correlated with clinical outcomes, morphological characteristics previously evaluated in whole slide tissue sections (hypercellularity/hypocellularity, round-oval or spindle dominant constituent cell (DCC) morphology, and necrosis), Ki67, overall survival, and RSS. Only one of the fifty-two cases studied showed progression. In the multivariate analysis, neither the presence nor absence of immune cells (B-lymphocytes, T-lymphocytes, and macrophages) showed any association with the assessed RSSs (Demicco, Sugita, G-score, and Huang). Interestingly, the case that showed progression had high immune infiltrate with expression of CD68, CD163, CD8, and CD20 markers (score of 3). Round-oval cell morphology was associated with the presence of higher levels of CD163 macrophages. Lastly, the scant presence of CD20+ lymphocytes correlated with less necrosis, and cases with higher PDL-1 expression correlated with increased Ki67 values. All cases were negative for LAG-1 and PD-1. SFT ITME components correlated with independent variables with prognostic significance. Nevertheless, ITME did not correlate with RSS scores.

Lymphomatoid Papulosis Type D in a Mestizo-Ancestry Man.

Lymphomatoid papulosis (LyP) belongs to the CD30+ skin lymphoproliferative disorders; it is defined as a chronic, recurrent, self-healing eruption of papules and small nodules with the histopathologic features of a cutaneous T-cell lymphoma. It is classified according to histopathology into subtypes A to F and with chromosomal rearrangement 6p25.3. Type D is characterized by epidermotropism of atypical CD8+ and CD30+ lymphocytes, small to medium size, forming papules and nodules with erosion and necrosis, which represents a formidable challenge in the differential diagnosis with aggressive cutaneous cytotoxic lymphomas. We present the clinical case of a 22-year-old man with subacute dermatosis, who underwent a skin biopsy with a report of LyP. Immunohistochemistry showed negative CD4, CD5, granzyme-B markers and positive CD3, CD30, CD8, CD56, and (T-cell intracellular antigen 1) TIA-1 markers, concluding the diagnosis of type D LyP. The course of the disease is recurrent; however, the prognosis is good with a 10-year survival of 100%. We present the case of a mestizo-ancestry patient who developed a type-D LyP, and, to the best of our knowledge, there are no publications of type D LyP from Latin-American authors or about mestizo-ancestry (or hispanic) patients; therefore, we consider of relevance to inform about these findings.

Longitudinal analysis of peripheral immune cells in patients with multiple sclerosis treated with anti-CD20 therapy.

Anti-CD20 therapy is a highly effective treatment for multiple sclerosis (MS). In this study, we investigated MS-related changes in peripheral blood mononuclear cell (PBMC) subsets compared to healthy controls and longitudinal changes related to the treatment. Multicolor spectral flow cytometry analysis was performed on 78 samples to characterize disease- and treatment-related PBMC clusters. Blood samples from MS patients were collected at baseline and up to 8 months post-treatment, with three collection points after treatment initiation. Unsupervised clustering tools and manual gating were applied to identify subclusters of interest and quantify changes. B cells were depleted from the periphery after anti-CD20 treatment as expected, and we observed an isolated acute, transitory drop in the proportion of natural killer (NK) and NKTcells among the main populations of PBMC (P = 0.03, P = 0.004). Major affected PBMC subpopulations were cytotoxic immune cells (NK, NKT, and CD8+ Tcells), and we observed a higher proportion of cytotoxic cells with reduced brain-homing ability and a higher regulatory function as a long-term anti-CD20-related effect. Additionally, anti-CD20 therapy altered distributions of memory CD8+ Tcells and reduced exhaustion markers in both CD4+ and CD8+ Tcells. The findings of this study elucidate phenotypic clusters of NK and CD8+ Tcells, which have previously been underexplored in the context of anti-CD20 therapy. Phenotypic modifications towards a more regulatory and controlled phenotype suggest that these subpopulations may play a critical and previously unrecognized role in mediating the therapeutic efficacy of anti-CD20 treatments.

EP110 - Chyloperitoneum in Follicular lymphoma – presenting as an inguinal hernia

Abstract Introduction Follicular lymphoma is one of the subtypes of slow growing Non-Hodgkin Lymphoma. Rarely, follicular lymphoma can present with chyloperitoneum or chylous ascites which is characterized by a milky-white coloured fluid with raised triglyceride levels. Case Report We present a case of a 60-year-old diabetic male, complaining of a right sided inguinal swelling for 1 year. Examination revealed a reducible right inguinal swelling with a positive cough impulse. The laboratory workup was unremarkable. Ultrasound scan of anterior abdominal wall showed a defect of 2.1 cm in right inguinal region with protrusion of abdominal contents and a positive cough impulse. A diagnosis of right sided reducible direct inguinal hernia was made. Patient was scheduled for Lichtenstein repair. Per-operatively, the hernial sac contained healthy omentum with milky-white ascitic fluid. Omentum was reduced and suture repair was performed. A drain was placed intra-peritoneally. The postoperative course of the patient was uneventful. Ascitic fluid analysis showed high triglyceride levels and no evidence of malignant cells, suggesting chylous ascites. Contrast enhanced CT scan of abdomen and pelvis revealed a large homogenous minimally enhancing soft tissue density retroperitoneal mass in peri aortic and paravertebral location with abdominal lymphadenopathy suggesting retroperitoneal lymphoma. Excisional biopsy of an enlarged inguinal lymph node confirmed follicular lymphoma with positive BCL2 and CD10 markers. Conclusion Follicular lymphoma presents in a variety of ways including chylous ascites in an inguinal hernia and a high index of suspicion is required to diagnose it.

Expression of antigen-presenting cells in lung of postmortem SARS-CoV-2 cases.

Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is a deadly pulmonary disease with impaired immunological response that causes significant tissue damage and organ failure. Postmortem examination of the lung is a useful tool for understanding the immunopathogenesis of this virus. Lung autopsy samples from seven dead SARS-CoV-2 patients were obtained and evaluated using hematoxylin and eosin stain to analyze the histopathological changes in those samples, on the other hand, Immunohistochemical (IHC) staining was used for detection of CD21, CD1a, CR1 (CD35), CD68, Myeloperoxidase (MPO), CD15, CD56, CD3, CD20, CD4, and CD8 cells markers. Histopathological examination revealed diffuse alveolar damage with extensive parenchymal architecture distortion, intravascular fibrin clot, deposition of collagen fibers, vascular congestions and blood vessels containing thrombi, pneumocyte type II with inflammatory cell infiltration. The IHC staining for the innate immune cells such as antigen-presenting cells (APCs) including dendritic cells, Macrophages, and neutrophils showed a strong positive staining, while CD56 Natural killer (NK) cells showed negative staining. On the other hand, the specific immune cells including; CD20 B cells, CD3 T cells, and CD4 helper T cells, showed positive staining while CD8 Cytotoxic T cells showed negative staining. The lung autopsy samples from patients with COVID-19 confirmed the presence of APCs through the positive staining of CD21, CD1a, CD35, CD68, MPO, and CD15 expressed the virus recognition, proinflammatory cytokine production, and adaptive immune cells activation through CD3, CD4, and CD20 positive staining and the role of APCs in the severity of pulmonary infection and pathogenesis of SARS-CoV-2 infection however the absence of the CD56 NK and CD8 cytotoxic T explains the worse infection status for the patients.

Identifying ADGRG1 as a specific marker for tumor-reactive T cells in acute myeloid leukemia

Besides chemotherapy and hematopoietic stem cell transplantation (HSCT), autologous T cells can also serve as a new treatment approach for AML patients. However, the features of tumor-reactive T cells and their distinctive markers still lack full description. To evaluate the characteristics of tumor-reactive T cells, we collected bone marrow (BM) T cells from newly diagnosed AML patients with RUNX1::RUNX1T1 as examples for paired single-cell RNA sequencing and single-cell V(D)J sequencing. Based on the STARTRAC-like algorithm, we defined bystander T cells and tumor-reactive T cells. Compared with bystander T cells, tumor-reactive T cells presented as senescent-like cytotoxic terminally differentiated T cells (Temra) with upregulated NK-related markers. Additionally, we found ADGRG1 could serve as the specific marker of CD8+ T tumor-reactive T cell and validated it through the Runx1Runx1t1/+; Mx1-Cre mouse model. In chimeric antigen receptor (CAR)-T and target cell system, ADGRG1 was selectively upregulated upon antigen-TCR encounter. Moreover, ADGRG1+CD8+ T cells released a higher level of IFN-γ and showed higher cell-killing ability when exposed to matched AML blasts. Together, our findings depict the single-cell profile of tumor-reactive T cells in AML BM and propose that ADGRG1 can act as an indicator of T cell tumor reactivity in AML, which may be further harnessed for adoptive cell therapy and tumor-reactive TCR enrichment.

Study of histopathological and ultrastructural changes in the lungs of COVID-19 patients

BackgroundCOVID 19 infection is a multi-systemic disease with the lungs bearing the maximum brunt. Very few autopsy studies have been done on the deceased patients. The present study aimed to study the clinical spectrum and lab parameters of COVID-19 patients along with the morphological spectrum of the lung changes in these patients by histology, Immunohistochemistry and Electron microscopy. We also compared the Clinical severity and Lab parameters with the histopathological phase of the lung involvement. MethodsWe conducted a cross sectional observational study between Jun 2021 to Dec 2022 where in needle necropsy of lung tissue of COVID 19 confirmed fatal cases were studied by histopathology, immunohistochemistry (using CD3, CD4, CD8, CD19, CD20, CD68 and Pan CK markers) and Electron microscopy. Various hematological, histopathological and ultrastructural parameters were studied. The results were analyzed using SPSS 25.0 software. ResultsA total of 24 cases were studied. The mean age was 65.9 yrs, male: female ratio was 2:1. Five had moderately severe COVID while 19 had severe COVID infection. Diffuse alveolar damage (DAD) was seen in 83% of all the cases. On the basis of clinical severity and histopathological staging, broadly two histopathological groups were made (Proliferative and Organizing). Difference between TLC, N/L ratio and L/Platelet ratio in two histopathological groups was assessed using unpaired t test, however no significant difference was noted. We did not find any correlation between clinical severity and histopathological groups though severe cases of COVID-19 were found to have a shorter time between admission to the hospital and death. All the biopsies were subjected to electron microscopy and in two of the cases we could demonstrate corona virus particles. ConclusionOur study has shown that severe cases of COVID 19 are associated with a shorter time between admission to hospital and death. While the histopathology of the lung showed diffuse alveolar damage as the most common finding; the Transmission Electron Microscopy (TEM) played an important role in characterizing the ultrastructure of these cells and demonstrating corona virus particles.

Bone Marrow and Peripheral Blood Mononuclear Cell Phenotype Changes after Cultivation and Autologous Infusion in Patients with Primary Biliary Cholangitis.

Primary biliary cholangitis (PBC) is a condition affecting the liver and immune system. In this study, the impact of autologous bone marrow-derived mononuclear cell (BM-MNC) transplantation on PBC patients was investigated. Sixteen eligible PBC patients participated at the National Scientific Medical Center in Astana, Kazakhstan, between 2017 and 2022, and BM-MNCs were harvested from their anterior iliac crest. After isolating and cultivating the BM-MNCs, they were infused back into the patient's peripheral veins. Changes in BM-MNC and peripheral blood mononuclear cell (PB-MNC) phenotypes were assessed before and after a 24-hour cultivation period and 72 hours post-transplantation. We monitored liver function parameters over 6-month intervals and conducted flow cytometry analysis to assess CD markers on BM-MNCs before and after cultivation and PB-MNCs before and after transplantation. Statistical analysis included the Friedman test for liver parameters and the Wilcoxon signed-rank test for BM-MNC and PB-MNC comparisons. Our findings revealed significant reductions in liver function tests after multiple transplantations. Flow cytometry analysis before and after a 24-hour culture and autologous BM-MNC infusion revealed the expansion of specific cell populations, with significant increases in CD3+, CD4+, CD16+, CD20+, CD25+, CD34+, CD105+, CD73+, СD117+, and CD34+populations, while CD4+25+, CD34+105+, and CD4+FOXP3+ populations decreased. Interestingly, a contradictory finding was observed with a decrease in bone marrow CD34+105+ cell lines (P=0.03) alongside an increase in peripheral CD34+105+ population (P=0.03). In summary, our study shows that BM-MNC transplantation in PBC patients leads to changes in immune cell populations and liver function. These findings suggest potential therapeutic applications of BM-MNC transplantation in managing PBC and offer insights into the dynamics of immune cells associated with this treatment approach.