Direct ex vivo flow cytometric analysis of human microglial cell CD4 expression: examination of central nervous system biopsy specimens from HIV-seropositive patients and patients with other neurological disease

被引:61
作者
Dick, AD
Pell, M
Brew, BJ
Foulcher, E
Sedgwick, JD
机构
[1] ROYAL PRINCE ALFRED HOSP,CENTENARY INST CANC MED & CELL BIOL,IMMUNOPATHOL RES GRP,SYDNEY,NSW 2042,AUSTRALIA
[2] ST VINCENTS HOSP,DEPT NEUROSURG,SYDNEY,NSW 2010,AUSTRALIA
[3] ST VINCENTS HOSP,DEPT NEUROL,SYDNEY,NSW 2010,AUSTRALIA
[4] ST VINCENTS HOSP,CTR IMMUNOL,SYDNEY,NSW 2010,AUSTRALIA
关键词
CD4; microglia; HIV; neurological/brain; AIDS dementia complex; FACS; flow cytometry;
D O I
10.1097/00002030-199714000-00006
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Objective: To define a clear ex vivo flow cytometric phenotype for adult human microglia that would distinguish it from all other macrophage lineage cells in the central nervous system (CNS) or blood, and to utilize this phenotype to examine the activation state and CD4 expression of microglia freshly derived from CNS tissue of HIV-positive patients and those with other neurological diseases. Design: Fresh human CNS tissue from both HIV-uninfected and HIV-infected individuals was obtained by biopsy or resection, and cells isolated immediately, labelled for flow cytometry and analysed. Methods: A Percoll density gradient isolation technique and phenotypic characteristics used for rodent microglia were applied and modified. Results: Resident microglia could clearly be defined by the flow cytometric phenotype CD45(low) CD4-CD11b+ CD11c(high) major histocompatibility complex (MHC) class II+ CD26-CD14-. Assuming normally low-level MHC class II expression in the healthy CNS, it was likely that MHC class II positivity reflected underlying pathology necessitating biopsy or resection and appeared to be a 'leaky' activation marker. Microglia activation was observed in specimens from only six (35%) out of 17 HIV-uninfected but all four (100%) HIV-infected patients, defined strictly as any level of upregulation of CD4 expression, to produce the phenotype CD45(low/medium) CD4(low) CD11b+CD11c(high) MHC class II+/++ CD26-CD14-. Where examined by immunohistology, CD68 was also upregulated in these cases. Conclusions: When activated in situ, microglia express low levels of CD4 and this is always seen in tissue from HIV-infected patients. Using the flow cytometric phenotype established here, microglia from HIV-infected tissue can now be isolated in pure form and studied directly ex vivo.
引用
收藏
页码:1699 / 1708
页数:10
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