Summary A mouse IgGi monoclonal antibody (1C4), which recognizes a cell surface molecule on murine natural cytotoxic{NC) cells was produced. By flowcytometry. IC4preferentially reacted with less than 5% of fresh CBA spleen cells and 20-50% of CBA-interleukin-3 (lL-3) cells, an in vitro derived NC-like cell line. In vitro treatment of spleen cells from a number of inbred mouse strains either with 1C4 alone or 1C4 coupled to dynabeads markedly decreased or abolished NC activity of the cells against ^'Cr-labelled WEHl-164 targets. Splenic NC activity of these same mouse strains was also reduced or abolished by in vivo administration of IC4. The effect was evident within 2 h of treatment and persisted for at least I week. In contrast IC4 had little or no effect on splenic NK activity against "Cr-labeiled YAC-I targets over the same range of experiments in vitro and in vivo. Results of strain surveys for both in vitro and in vivo reduction of splenic NC activity by 1C4 treatment showed that CBA. C57BL/6, BALB/c and NZB mice were positive and CE and DBA/2 mice were negative, indicating that 1C4 recognizes an allo-antigen on mouse NC cells. This allo-antibody has been designated NC-II, and thus IC4 is an anti-NC-11 monoclonal antibody.
SummaryAs part of the strategy for screening for natural kilter (NK) cell-specific monoclonal antibodies (MoAb) we have raised a number of murine NK-like cell lines in media containing interleukin-2 (IL-2). The detection of specific NK cell alloantigens on a C57BL/6 cell line in long-term culture in IL-2 is the subject of this paper. The C57BL/6 cell line has the morphology of large granular lymphocytes (LGL) and exhibits strong cytolytic activity against the archetype NK cell target, YAC-I. Absorption of three anti-NK antiserum. NZB anti-BALB/c (anti-NK-2-1), BALB/c anti-DBA/2 {anti-NK-31) and CE anti-CBA (anti-NK-4 I), with the C57BL/6 cell line removed the anti-NK activity from these antisera. Flow cytometric studies ofthe C57BL/6 cell line demonstrated significant binding ofthe anti-NK-1-1 MoAb produced hy hybridoma PK136. Our results suggest that the C57BL/6 NK-like cell line exhibits some ofthe properties of naive NK cells and expresses all the known NK cell-specific alloantigens, NK-1 -1, NK-2-1. NK-3-1 and NK-4' I and therefore is potentially useful in selecting NK specific hybridomas and in studying the biology of NK cells.
Conventional techniques used for the analysis of antisperm antibodies are not suited to the mouse model because of their requirements for relatively large amounts of serum and their inability to handle large numbers of samples. This has inhibited use of the mouse as an experimental model in areas involving antisperm immunity. As the ELISA technique has been successfully applied to analysis of antisperm antibody in human sera, we investigated its use as an assay for screening antisperm antibody in mouse serum. This report describes a simplified version of the ELISA technique that we have found to be successful for this purpose. The assay described can assess levels and classes of antisperm antibody in mouse serum and can also be used as a screening assay for monoclonal antibodies to mouse sperm. It should facilitate use of the mouse in experimental work in areas involving assessment of immunity to sperm.
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