T cells direct macrophage recruitment and giant cell formation but not proliferation in experimental crescentic glomerulonephritis

SUMMARY: The role of T cells in directing macrophage recruitment and proliferation and giant cell formation was studied in crescentic glomerulonephritis (GN) in rats. Development of GN was associated with a significant glomerular influx of T cells and macrophages and crescent formation. Multi-nucleated giant cells (MGCs) were also prominent in glomeruli and most frequently associated with crescents. Their phenotype suggested that they were derived from recently recruited blood monocytes. the majority of glomerular macrophages and MGCs showed evidence of recent proliferation. T cell depletion using anti-CD5 or anti-CD4 monoclonal antibodies significantly reduced T cells and macrophage accumulation, crescent formation (anti-CD5 treated 12 ± 4.0%; anti-CD4 treated 13 ± 3.0%; control 64 ± 10%; P <0.001) and MGC formation [anti-CD5 treated 0.06 ± 0.01 cells per glomerular cross section (c/gcs); anti-CD4 treated 0.06 ± 0.01 c/gcs; control treated 1.2 ± 0.1 c/gcs; P <0.001]. the proliferation index (fraction of cells expressing proliferating cell nuclear antigen) of macrophages and MGCs was not affected by T cell depletion (macrophage proliferation index: anti-CD5 treated 0.88 ± 0.04; anti-CD4 treated 0.83 ± 0.02; control 0.85 ± 0.02; MGC proliferation index: anti-CD5 treated 0.84 ± 0.06; anti-CD4 treated 0.86 ± 0.04; control 0.90 ± 0.05). These studies demonstrate that glomerular macrophage recruitment, MGC and crescent formation are dependent on T helper cells but proliferation of macrophages and MGCs is independent of T cells. This suggests that the major mechanism for T cell induced macrophage accumulation in crescentic GN is via directing their recruitment rather than their local proliferation. Giant cell formation, which is also predominantly T cell dependent, appears to be independent of their proliferative activity.     

人类重组肝细胞生长因子影响膀胱癌细胞株生长和抑制丝裂霉素C诱导细胞凋亡的研究

目的 :观察人类重组肝细胞生长因子 (rhHGF)对膀胱移行细胞癌T2 4细胞增殖和丝裂霉素C(MMC)诱导的细胞凋亡的影响。方法 :以T2 4细胞为研究材料 ,用不同浓度rhHGF干预T2 4细胞生长及MMC诱导的细胞凋亡作用。利用MTT实验、流式细胞仪技术来判定rhHGF对细胞增殖和凋亡的影响。结果 :不同浓度rhHGF组间 ,T2 4相对增殖细胞数差异无统计学意义 (P >0 .0 5 ) ;rhHGF预处理后 ,MMC诱导的T2 4细胞凋亡率明显下降 ,在一定范围内有量效关系。结论 :HGF并不显著诱导T2 4细胞增殖 ,而一定剂量范围内的HGF能抑制MMC诱导的细胞凋亡 ,可能与膀胱癌的复发、耐药等临床特性有关     

Mucosal associated invariant T cells are differentially impaired in tolerant and immunosuppressed liver transplant recipients

Mucosal associated invariant T (MAIT-) cells represent a semi-invariant T cell population responsive to microbial vitamin B metabolite and innate cytokine stimulation, executing border tissue protection and particularly contributing to human liver immunity. The impact of immunosuppressants on MAIT cell biology alone and in context with solid organ transplantation has not been thoroughly examined. Here, we demonstrate that in vitro cytokine activation of peripheral MAIT cells from healthy individuals was impaired by glucocorticoids, whereas antigen-specific stimulation was additionally sensitive to calcineurin inhibitors. In liver transplant (LTx) recipients, significant depletion of peripheral MAIT cells was observed that was largely independent of the type and dosage of immunosuppression, equally applied to tolerant patients, and was reproducible in kidney transplant recipients. However, MAIT cells from tolerant LTx patients exhibited a markedly diminished ex vivo activation signature, associated with individual regain of functional competence toward antigenic and cytokine stimulation. Still, MAIT cells from tolerant and treated liver recipients exhibited high levels of PD1, accompanied by functional impairment particularly toward bacterial stimulation that also affected polyfunctionality. Our data suggest interlinked effects of primary liver pathology and immunosuppressive treatment on overall MAIT cell fitness after transplantation and propose their monitoring in context with tolerance induction protocols.     

十色流式检测HIV/AIDS患者外周血中T淋巴细胞亚群及活化状态

目的建立十色流式方案检测HIV/AIDS患者外周血中T淋巴细胞亚群及活化状态。 方法按多色流式配色原则,初步确立抗人CD3、CD4、CD8、CD45RA、CD25、CCR7、CD28、CD38、HLA-DR抗体及7-AAD的配色方案。采用1例HIV/AIDS患者外周血PBMC标本进行最适电压调节、荧光补偿设定和FMO对照;设定条件后,检测5例HIV/AIDS患者的外周血样本。 结果建立检测人外周血中T淋巴细胞亚群及活化状态的最优十色流式染色方案,采用此方案检测5例HIV/AIDS患者外周血标本,分析T淋巴细胞各亚群比例及活化程度,发现不同细胞亚群中活化分子HLA-DR、CD38、CD28的表达模式存在显著差异。 结论十色流式方案可检测HIV/AIDS患者外周血中T淋巴细胞亚群及其活化状态操作简便,结果可靠。     

Myeloid‐derived suppressor cells increase and inhibit donor‐reactive T cell responses to graft intestinal epithelium in intestinal transplant patients

Recent advances in immunosuppressive regimens have decreased acute cellular rejection (ACR) rates and improved intestinal and multivisceral transplant (ITx) recipient survival. We investigated the role of myeloid‐derived suppressor cells (MDSCs) in ITx. We identified MDSCs as CD33⁺CD11b⁺ lineage(CD3/CD56/CD19)^?HLA‐DR^?/low cells with 3 subsets, CD14^?CD15^? (e‐MDSCs), CD14⁺CD15^? (M‐MDSCs), and CD14^?CD15⁺ (PMN‐MDSCs), in peripheral blood mononuclear cells (PBMCs) and mononuclear cells in the grafted intestinal mucosa. Total MDSC numbers increased in PBMCs after ITx; among MDSC subsets, M‐MDSC numbers were maintained at a high level after 2 months post ITx. The MDSC numbers decreased in ITx recipients with ACR. MDSC numbers were positively correlated with serum interleukin (IL)‐6 levels and the glucocorticoid administration index. IL‐6 and methylprednisolone enhanced the differentiation of bone marrow cells to MDSCs in vitro. M‐MDSCs and e‐MDSCs expressed CCR1, ‐2, and ‐3; e‐MDSCs and PMN‐MDSCs expressed CXCR2; and intestinal grafts expressed the corresponding chemokine ligands after ITx. Of note, the percentage of MDSCs among intestinal mucosal CD45⁺ cells increased after ITx. A novel in vitro assay demonstrated that MDSCs suppressed donor‐reactive T cell–mediated destruction of donor intestinal epithelial organoids. Taken together, our results suggest that MDSCs accumulate in the recipient PBMCs and the grafted intestinal mucosa in ITx, and may regulate ACR.     

Predictive value of laminin-5 and membrane type 1-matrix metalloproteinase expression for cervical lymph node metastasis in T1 and T2 squamous cell carcinomas of the tongue and floor of the mouth

BACKGROUND: Laminin-5 (Ln-5) cleaved by membrane type 1-matrix metalloproteinase (MT1-MMP) enhances the migration of tumor cells. The purpose of this study was to determine whether or not enhanced expression of both Ln-5 and MT1-MMP was associated with lymph node metastasis in patients with T1 and T2 squamous cell carcinoma (SCC) of the tongue and floor of the mouth. METHODS: By use of biopsy specimens of primary tumors from 57 patients, intratumoral expression of Ln-5 and MT1-MMP was evaluated immunohistochemically and its association with node metastasis analyzed. RESULTS: The tumors were categorized into three groups: Ln-5 focal type/MT1-MMP (-) (group I, n = 14), Ln-5 focal type/ MT1-MMP (+) and Ln-5 diffuse type/MT1-MMP (-) (group II, n = 16), and Ln-5 diffuse type/MT1-MMP (+) (group III, n = 27). The incidence of node metastasis (initial and latent metastases) was two of 14 (14.3%), five of 16 (31.3%), and 15 of 27 (55.6%) in groups I, II, and III, respectively. Multivariate analysis identified tumor thickness (odds ratio, 4.751; p = .0152) and Ln-5/ MT1-MMP expression (odds ratio, 3.795, p = .0304) as independent factors of node metastasis. Moreover, in 35 patients with N0 disease, Ln-5/MT1-MMP expression was the only parameter associated with latent node metastasis (odds ratio, 12.800, p = .0247). CONCLUSION: These results suggest that immunohistochemical evaluation of Ln-5 and MT1-MMP expression is useful for identifying patients with T1 and T2 SCC of the tongue and floor of the mouth who should be treated with elective neck dissection.