Abstract:
:Ammonium-chloride-containing solutions (AC) are routinely used to lyse red blood cells during preparation of PBMC. Although exposure to AC has been described to affect the ultrastructural appearance of large granular lymphocytes and to temporarily inhibit cytolytic activity of PBMC preparations, the cellular basis of this phenomenon has not been studied. Here, the inhibitory effect of AC on human CTL and NK-mediated cytotoxicity has been analyzed in 4-h 51Cr-release assays. The results show that NK killing of K562 leukemia cells and xenogeneic endothelial cells is inhibited by AC exposure. The effect is dose-dependent and reversible, because recovery of cytotoxicity is observed within 15 h of re-culturing. AC does not reduce the viability of NK cells and the inhibitory effect is not mediated by the exhaustive release of granzymes upon AC treatment. In contrast, antigen-specific CTL killing of EBV-transformed B-lymphoblastoid cell lines and xenogeneic PHA lymphoblasts was less sensitive to AC and data are presented suggesting that FasL-induced apoptosis is not inhibited by AC. In conclusion, perforin-mediated NK killing is AC-sensitive whereas CTL killing and FasL-mediated killing appear to be AC-resistant. Therefore, AC represents a powerful tool to study different mechanisms of cell-mediated cytotoxicity and may be helpful in assessing antigen-specific CTL cytotoxicity without the influence of NK cell-mediated background killing.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Brander C,Matter-Reissmann UB,Jones NG,Walker BD,Sachs DH,Seebach JDdoi
10.1016/s0022-1759(01)00326-xkeywords:
subject
Has Abstractpub_date
2001-06-01 00:00:00pages
1-14issue
1-2eissn
0022-1759issn
1872-7905pii
S002217590100326Xjournal_volume
252pub_type
杂志文章abstract::Structural point mutations in exon 1 at codons 52, 54 and 57 and a promotor polymorphism at -221 bp of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to various infectious diseases. We developed a genotyping method based on the 5' nuclease (TaqMan) assay in combination with the use ...
journal_title:Journal of immunological methods
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pub_type: 杂志文章
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journal_title:Journal of immunological methods
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pub_type: 临床试验,杂志文章,多中心研究
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journal_title:Journal of immunological methods
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pub_type: 杂志文章
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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pub_type: 杂志文章
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