Lung cancer is the leading cause of cancer mortality among people with HIV (PWH), with increased incidence and poor outcomes. This study explored whether the tumor microenvironment (TME) of HIV-associated non-small cell lung cancer (NSCLC) limits tumor-specific immune responses. With a matched cohort of NSCLC from PWH and people without HIV (PWOH), we used imaging mass cytometry, linear mixed effects model and AI-based pageRank mathematical algorithm based on spectral graph theory to demonstrate that HIV-associated tumors demonstrate differential distribution of tumor infiltrating CD8+ and CD4+ T cells, enriched for the expression of PD-1 and Lag-3, as well as activation and proliferation markers. We also demonstrate higher expression of immunoregulatory molecules (PD-L1, PD-L2, B7-H3, B7-H4, IDO1 and VISTA), among tumor-associated macrophages. Discrimination of cells between tumors from PWH versus PWOH was confirmed by spectral graph theory with 84.6% accuracy. Furthermore, we noted differences in spatial orientation of immune cells within the TME of PWH compared to PWOH. Additionally, cells from PWH, compared to PWOH, exhibited decreased tumor killing when exposed to HLA-matched NSCLC cell lines. In conclusion, our study demonstrates that the HIV-associated tumor microenvironment sustains a unique immune landscape, with evidence of immune cells with enhanced immunoregulatory phenotypes and impaired anti-tumor responses, with implications for response to immune checkpoint blocker therapies.

Chronic idiopathic thrombocytopenic purpura (ITP) is an autoimmune disease characterized by a breakdown of immune tolerance; in ITP, the body's immune system mistakenly attacks and destroys platelets. This study aims to investigate the role and underlying mechanisms of FOXP3 in chronic ITP.
Flow cytometry was used to detect the proportion of CD4+CD25+FOXP3+ regulatory T cells (Tregs) in CD4+CD25+ T lymphocytes from 20 patients with chronic ITP (CITP), 20 acute ITP (AITP) controls, and 20 healthy individuals.CD4+CD25+ Treg cells were isolated from peripheral blood of patients with CITP using magnetic beads and then treated with phosphate-buffered saline solution or decitabine (a methylation inhibitor) for 48 h. The levels of interleukin-2 (IL-2), IL-10, and transforming growth factor-beta1 (TGF-β1) in the plasma and CD4+CD25+ Treg cells were assessed by Enzyme-linked-immunosorbent serologic assay and quantitative real-time polymerase chain reaction (qRT-PCR). FOXP3 level was measured by qRT-PCR and Western blot analysis. Methylation-specific PCR (MS-PCR) was adopted to detect the status of FOXP3 methylation.
The number of Treg cells and the contents of IL-2, IL-10, and TGF-β1 decreased in patients with CITP, compared to the AITP control group and normal group. FOXP3 expression was reduced and FOXP3 methylation increased in patients with CITP, compared to the AITP control group and normal group. Hypermethylation of FOXP3 promoter led to decrease in FOXP3 level in Treg cells. Inhibition of FOXP3 promoter hypermethylation promoted the secretion of IL-2, IL-10, and TGF-β1 in Treg cells.
The number of Treg cells in CITP patients decreased, and the hypermethylation of FOXP3 promoter led to reduction of its expression in Treg cells, thus affecting the immune functioning of Treg cells.

Background Among people with HIV (PWH), lung cancer is the leader cause of cancer mortality, with increased risk and poor clinical outcomes compared to people without HIV (PWOH). HIV is known to result in persistent global immune dysfunction despite antiretroviral therapy, but little is known about the lung cancer tumor microenvironment. This study explored whether the tumor microenvironment (TME) of HIV-associated non-small cell lung cancer (NSCLC) is associated with an immunoregulator environment that limits tumor-specific immune responses. Methods A tissue microarray was constructed with NSCLC tumors from 18 PWH and 19 PWOH (matched for histological subtype, stage, year of diagnosis, age, sex and smoking status), and incubated with metal-conjugated antibodies for evaluation by imaging mass cytometry (IMC). IMC marker scores were extracted by automated cell segmentation and single-cell data was analyzed by Phenograph using unsupervised cell-segmentation and clustering of cells. Evaluation of tumor infiltrating immune cells, CD4+ and CD8+ T cells as well as CD68+ tumor associated macrophages were characterized for marker expression using a linear mixed-effects model. Additionally, a computational strategy based on the PageRank mathematical algorithm was used in order to establish an unsupervised and cell segmentation-independent signature associated with HIV status to discriminate differential expression of immune cell markers within the TME of the two groups. Peripheral blood mononuclear cells (PBMCs) from HLA-A02 donors (PWH and PWOH) were co-incubated with HLA-A02 lung cancer cell lines to quantify tumor killing (by Annexin V staining) and expression of T cell markers Lag-3 and CD25. Results Within the TME from HIV+ tumors, there is comparable level of infiltration of lymphocytes and tumor associated macrophages (TAMs) compared to non-HIV tumors, with a trend towards increased CD8+ T cells and decreased CD4:CD8 ratio among HIV+ tumors. Using a random effects model of individual markers, HIV+ tumors revealed increased expression of Ki67 and Granzyme B (GRZB) among CD8+ T cells; increased Ki67 and PD-1 among CD4+ T cells; and increased PD-L1, PD-L2, and Ki67 among TAMS. Unsupervised clustering analysis from IMC data demonstrated differential distribution of tumor infiltrating CD8+ T cell clusters between HIV+ and non-HIV tumors, defined by marker expression patterns. Three clusters were significantly elevated in HIV+ tumors (57.1% vs. 21.7% in non-HIV tumors, p<0.0001). All three clusters had comparatively elevated PD-1 and Lag-3 expression with varying expression of activation and proliferation markers CD25 and Ki67. Within tumor-infiltrating CD4+ T cells, a cluster characterized by checkpoint protein expression (PD-1+ and LAG-3) was also highly represented in HIV+ cases (35.2% vs. 9.8% in non-HIV cases, p<0.0001). HIV+ tumor-associated macrophages (TAM) had higher expression of immunoregulatory molecules (PD-L1, PD-L2, B7-H3, B7-H4, IDO1 and VISTA), confirmed by the expansion of three clusters comprising 58.8% of TAMs vs. 17.8% in non-HIV tumors (p<0.0001). Discrimination of cells between HIV+ and HIV-TME was further confirmed by spectral graph theory with 84.6% accuracy, with a combination of markers on TAMs and T cells. Lastly, PBMCs from PWH exhibited decreased tumor killing when exposed to HLA-matched NSCLC cell lines compared to PBMCs from PWOH. CD8+ T cells from PWH additionally had increased expression of immune checkpoint inhibitor Lag-3 upon exposure to tumor cells. Conclusions Our study demonstrates that the TME of HIV+ patients is characterized by a unique immune landscape, distinct from that of PWOH, with evidence of expansion of immune cells with enhanced immunoregulatory phenotypes and associated with impaired anti-tumor responses.

Herpes zoster (HZ) is caused by the varicella-zoster virus (VZV), and 20% of healthy humans and 50% of people with immune dysfunction have a high probability of suffering from HZ. This study aimed to screen dynamic immune signatures and explore the potential mechanism during HZ progression.
Peripheral blood samples from 31 HZ patients and 32 age-sex-matched healthy controls were collected and analyzed. The protein levels and gene levels of toll-like receptors (TLRs) were detected in peripheral blood mononuclear cells (PBMCs) by flow cytometry and quantitative real-time PCR. Further, the characteristics of T cell subsets and cytokines were detected via a cytometric bead array.
Compared to healthy controls, the mRNA levels of TLR2, TLR4, TLR7, and TLR9 mRNA in PBMCs were significantly increased in HZ patients. The protein level of TLR4 and TLR7 was significantly increased in HZ patients, but the levels of TLR2 and TLR9 were dramatically decreased. The CD3+ T cells were constant in HZ and healthy controls. CD4+ T cells were decreased in HZ patients, while CD8+ T cells were increased, resulting in an improved CD4+/CD8+ T cells ratio. Further, it was found that Th2 and Th17 were not changed, but the decreased Th1 and upregulated Treg cells were found in HZ. The Th1/Th2 and Th17/Treg ratios were significantly decreased. Last, the levels of IL-6, IL-10, and IFN-γ were significantly increased, but IL-2, IL-4, and IL-17A had no significant changes.
The dysfunction of host's lymphocytes and activation of TLRs in PBMCs were the important mechanism in varicella-zoster virus induced herpes zoster. TLRs might be the core targets for the therapy drug development in treating HZ.
© 2023 Chen et al.

Signaling via the OX40/OX40L axis plays a key role in CD4+ T cell development, and OX40L expression is primarily restricted to antigen-presenting cells (APCs). This study was designed to assess the role of APC-mediated OX40L expression in the context of the development of rheumatoid arthritis (RA)-associated CD4+ T cell subsets. For these analyses, clinical samples were harvested from patients with osteoarthritis and RA, with additional analyses performed using OX40-/- mice and mice harboring monocyte/macrophage-specific deletions of OX40L. Together, these analyses revealed tissue-specific roles for OX40/OX40L signaling in RA. Specifically, higher levels of synovial macrophage OX40L expression were associated with the enhanced development of T follicular helper cells in the joint microenvironment, thereby contributing to the pathogenesis of RA. This Tfh differentiation was found to be OX40/OX40L-dependent in this synovial setting. Overall, these results indicate that the expression of OX40L by synovia macrophages is necessary to support Tfh differentiation in the joint tissues, thus offering new insight regarding the etiological basis for RA progression.