Abstract
This study was performed to determine what factors related to the enzymatic isolation technique and assay conditions may influence the measurement of spontaneous cell-mediated cytotoxicity (SCMC) of mononuclear cells (MNC) isolated from human intestinal mucosa. In 18 studies, the SCMC of freshly isolated cells was 1.8 +/- 0.4% but increased to 12 +/- 3% following 24 hr culture without a change in the proportion of cells with the NK phenotype (Leu-7+). The SCMC tended to plateau with more prolonged culture. Culturing peripheral blood (PB) MNC for 24 hr did not alter SCMC nor the proportion of Leu-7+ cells. However, the suppression of the SCMC of PBMNC preincubated with the supernatant of the collagenase digestion of intestinal mucosa was completely reversed by 24 hr culture. Intestinal MNC were found to suppress the SCMC of autologous PBMNC in mixing experiments. However, 24 hr culture did not affect the degree of suppression and the proportion of T cells of the suppressor-cytotoxic phenotype (Leu-2a+) was also unchanged. It is concluded that the SCMC of intestinal MNC may be accurately assessed following 24 hr culture of the cells to allow recovery from the suppressive influences of the isolation process and that this does not introduce other artefactual problems. However, suppression of cytotoxicity within the assay may result in an underestimation of the SCMC of intestinal MNC when compared to that of PBMNC.
Full text
PDF







Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abo T., Balch C. M. A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1). J Immunol. 1981 Sep;127(3):1024–1029. [PubMed] [Google Scholar]
- Adams D. O., Snyderman R. Do macrophages destroy nascent tumors? J Natl Cancer Inst. 1979 Jun;62(6):1341–1345. [PubMed] [Google Scholar]
- Bull D. M., Bookman M. A. Isolation and functional characterization of human intestinal mucosal lymphoid cells. J Clin Invest. 1977 May;59(5):966–974. doi: 10.1172/JCI108719. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cerf-Bensussan N., Schneeberger E. E., Bhan A. K. Immunohistologic and immunoelectron microscopic characterization of the mucosal lymphocytes of human small intestine by the use of monoclonal antibodies. J Immunol. 1983 Jun;130(6):2615–2622. [PubMed] [Google Scholar]
- Chiba M., Bartnik W., ReMine S. G., Thayer W. R., Shorter R. G. Human colonic intraepithelial and lamina proprial lymphocytes: cytotoxicity in vitro and the potential effects of the isolation method on their functional properties. Gut. 1981 Mar;22(3):177–186. doi: 10.1136/gut.22.3.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clancy R. Isolation and kinetic characteristics of mucosal lymphocytes in Crohn's Disease. Gastroenterology. 1976 Feb;70(2):177–180. [PubMed] [Google Scholar]
- Falchuk Z. M., Barnhard E., Machado I. Human colonic mononuclear cells: studies of cytotoxic function. Gut. 1981 Apr;22(4):290–294. doi: 10.1136/gut.22.4.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibson P. R., Dow E. L., Selby W. S., Strickland R. G., Jewell D. P. Natural killer cells and spontaneous cell-mediated cytotoxicity in the human intestine. Clin Exp Immunol. 1984 May;56(2):438–444. [PMC free article] [PubMed] [Google Scholar]
- Gibson P. R., Verhaar H. J., Selby W. S., Jewell D. P. The mononuclear cells of human mesenteric blood, intestinal mucosa and mesenteric lymph nodes: compartmentalization of NK cells. Clin Exp Immunol. 1984 May;56(2):445–452. [PMC free article] [PubMed] [Google Scholar]
- Goodacre R., Davidson R., Singal D., Bienenstock J. Morphologic and functional characteristics of human intestinal lymphoid cells isolated by a mechanical technique. Gastroenterology. 1979 Feb;76(2):300–308. [PubMed] [Google Scholar]
- Kay H. D., Smith D. L. Regulation of human lymphocyte-mediated natural killer (NK) cell activity. I. Inhibition in vitro by peripheral blood granulocytes. J Immunol. 1983 Jan;130(1):475–483. [PubMed] [Google Scholar]
- Nair M. P., Schwartz S. A. Suppression of natural killer activity and antibody-dependent cellular cytotoxicity by cultured human lymphocytes. J Immunol. 1981 Jun;126(6):2221–2229. [PubMed] [Google Scholar]
- Ortaldo J. R., Bonnard G. D., Herberman R. B. Cytotoxic reactivity of human lymphocytes cultured in vitro. J Immunol. 1977 Oct;119(4):1351–1357. [PubMed] [Google Scholar]
- Pross H. F., Baines M. G. Studies of human natural killer cells. I. In vivo parameters affecting normal cytotoxic function. Int J Cancer. 1982 Apr 15;29(4):383–390. doi: 10.1002/ijc.2910290404. [DOI] [PubMed] [Google Scholar]
