Circulating CD3+ CD14+ T Cell-Monocyte complexes are dynamic, metabolically glucose-dependent HIV reservoirs and increased in individuals with glucose intolerance. Circulating CD3+ CD14+ T Cell-Monocyte complexes are dynamic, metabolically glucose-dependent HIV reservoirs and increased in individuals with glucose intolerance

Persistent and elevated systemic inflammation contributes to the increased risk of cardiovascular and metabolic diseases in persons with HIV (PWH). , but aspects of the innate and adaptive immune response to HIV and other chronic co-infections that promote the development of these comorbid conditions remain unclear. We used mass cytometry and the tracking responders expanding (T-REX) workflow to define differences in circulating cells of the innate and adaptive immune arms, which differentiate non-diabetic and prediabetic/diabetic persons with HIV (PWH) on long-term antiretroviral therapy (ART). We found a significantly higher proportion of circulating CD14+ monocytes complexed with T cells, B cells, and NK cells among PWH with diabetes. The CD3+ T cells in these complexes were predominantly CD8+ memory T cells. The CD3+ CD14+ T cell-monocyte complexes are pro-inflammatory and negatively associated with plasma interleukin-10 levels and circulating CD4+ T regulatory cells. Using transmission electron microscopy, we observed heterogeneous interactions between CD3+ T cells and monocytes within the complexes, including formation of immune synapses and phagocytosis. 10x Chromium single-cell sequencing RNA transcriptomic analysis of CD3+ CD14+ doublets showed differential expression of genes involved in T cell activation and the adaptive immune response compared to monocytes that were not complexed with other immune cells. Notably, there were significantly more copies of HIV RNA per cell in the CD3+ CD14+ T cell-monocyte complexes compared to CD14+ monocytes or CD3+ CD4+ T cells alone, and HIV virions were observed in both T cells and monocytes within the doublets?. Metabolically, CD3+ CD14+ T cell-monocyte complexes had high glucose-dependence with minimal mitochondrial dependence. Taken together, CD3+ CD14+ T cell-monocyte pairs are functional and physically interacting immune cell complexes which might be enriched with HIV. These complexesare increased among individuals with diabetes and may be a target for future HIV cure studies and diseases of aging. Overall design: Circulating peripheral blood mononuclear cells were isolated and analyzed using scRNAseq.

持续升高的全身性炎症会增加人类免疫缺陷病毒（HIV）感染者（PWH）罹患心血管疾病与代谢性疾病的风险，但目前针对HIV及其他慢性合并感染的先天与适应性免疫应答中，促进这类共病发生的相关机制仍不明确。本研究采用质谱流式细胞术（mass cytometry）与追踪应答扩增（T-REX）工作流，针对接受长期抗逆转录病毒治疗（ART）的HIV感染者，分析其先天及适应性免疫系统的循环细胞在非糖尿病、糖尿病前期与糖尿病患者间的差异。研究发现，在糖尿病HIV感染者体内，与T细胞、B细胞及自然杀伤细胞（NK cells）结合的循环CD14阳性单核细胞比例显著升高。这类复合物中的CD3阳性T细胞主要为CD8阳性记忆T细胞。CD3+CD14+ T细胞-单核细胞复合物具有促炎特性，且与血浆白细胞介素10（interleukin-10）水平及循环CD4阳性调节性T细胞数量呈负相关。通过透射电子显微镜观察，研究人员发现复合物内CD3阳性T细胞与单核细胞间存在多种异质性相互作用，包括免疫突触形成与吞噬作用。对CD3+CD14+双联体细胞进行10x Chromium单细胞RNA转录组分析显示，与未结合其他免疫细胞的单核细胞相比，这类复合物中参与T细胞活化及适应性免疫应答的基因存在差异表达。值得注意的是，CD3+CD14+ T细胞-单核细胞复合物内每细胞的HIV RNA拷贝数显著高于单独的CD14+单核细胞或CD3+CD4+ T细胞，且在双联体细胞的T细胞与单核细胞内均观察到HIV病毒粒子。代谢层面上，CD3+CD14+ T细胞-单核细胞复合物具有高度葡萄糖依赖性，线粒体依赖性极低。综上，CD3+CD14+ T细胞-单核细胞配对是具有功能且存在物理相互作用的免疫细胞复合物，且可能富集HIV病毒。这类复合物在糖尿病个体中数量增多，有望成为未来HIV治愈研究及衰老相关疾病的干预靶点。整体实验设计：分离外周血单个核细胞（PBMC）并采用单细胞RNA测序（scRNAseq）进行分析。