Endometrial lymphocyte fluctuations during the menstrual cycle may reflect hormonal regulation of maternal immunity, thereby promoting tolerance at the time of implantation.
No increase of intra-epithelial lymphocytes in the coeliac lesion may require us to reconsider the definition of coeliac disease as an inflammatory condition. Low CD4CD8 populations in treated as well as untreated coeliac patients indicate that these T cells are inherently absent in individuals genetically predisposed to coeliac disease.
Human endometrium is a rich source of lymphocytes which may have unique immunoregulatory functions. The aim of this study was to compare current procedures for endometrial tissue disaggregation, and optimise a method for isolation of endometrial lymphocytes. Tissue was obtained from 41 women undergoing elective hysterectomy or dilation and curettage (D&C) for reasons of benign non-endometrial pathology. Specimens were exposed to reduction/chelation, mechanical or enzymatic disruption. Optimal single cell suspensions of high yields (mean 8.8 x 10(6) range 3.5-18 x 10(6)lymphs) and good viability (60%) were obtained, using a combination of collagenase IV (200 U/ml) and DNase I (35 U/ml). Suspensions were further purified by density gradient centrifugation. Multi-colour flow cytometry was used for analysis of endometrial lymphocyte subsets. Cell suspensions were stained with mAbs specific for CD3, CD4, CD8, CD56, CD45 and CD14, and it was clearly shown that the developed method had no effect on surface glycoprotein expression. Phenotypic analysis revealed consistent populations of endometrial large granular lymphocytes (CD56+CD3-) 54.16%, and T-cells (CD3+) 37.73%. This technique was applicable to the characterisation of T-cell populations, including CD8+ (56.6%), CD4+ (44.0%), and particularly smaller populations of CD4+CD8+(3.56%), CD4-CD8-(3.34%) and CD56+(6.3%) due to it's sensitivity. In conclusion, optimised enzymatic digestion, in combination with flow cytometry provides an effective method for phenotypic examination of small endometrial lymphocyte subpopulations.
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