The role of CCL5 (RANTES) in the immune response against Mycobacterium tuberculosis in the guinea pig

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Title: The role of CCL5 (RANTES) in the immune response against Mycobacterium tuberculosis in the guinea pig
Author: Skwor, Troy Arthur
Abstract: Tuberculosis is the second leading cause of morbidity and mortality worldwide due to an infectious disease . Development of a new tuberculosis (TB ) vaccine would be facilitated by a better understanding of the mechanisms of protection induced by the current TB vaccine , Mycobacterium bovis BCG . Recombinant guinea pig (rgp )CCL5 and anti -rgpCCL5 were developed and characterized . The biological activity of rgpCCL5 was determined in a chemotaxis assay using T lymphocytes and pleural exudate cells . The specificity of rabbit anti -rgpCCL5 polyclonal antibody was confirmed by Western blot . RgpCCL5 was used to stimulate alveolar and peritoneal macrophages in vitro . and cytokine /chemokine gene expression was evaluated using real -time PCR . RgpCCL5 stimulated TNF & #945 ; , IL -1 & #946 ; , CCL2 , and CXCL8 mRNA expression and TNF & #945 ; protein production (as assessed in the L929 cell bioassay ) in macrophages . The effect of BCG -vaccination on CCL5 expression and production in leukocytes infected with M . tuberculosis was examined in vitro and in vivo . Polyclonal anti -rgpCCL5 was used to develop an ELISA assay to quantify gpCCL5 protein levels , and real -time PCR was used to detect CCL5 mRNA . Leukocytes isolated from BCG -vaccinated guinea pigs and infected in vitro with virulent M . tuberculosis demonstrated significantly elevated gpCCL5 mRNA and protein compared to cells from naive animals . The response of gpCCL5 to M . tuberculosis in vivo was studied in tuberculous pleural effusions , where peak levels of CCL5 mRNA and protein were reached at day 4 post -induction . Disease severity , cellular differentiation , histology , and cytokine /chemokine mRNA levels in pleural cells and granulomas were analyzed on day 4 in guinea pigs induced with tuberculous pleurisy and treated with either rgpCCL5 or anti -rgpCCL5 by direct intra -pleural injection . In these studies , neutralizing CCL5 resulted in reduced macrophage accumulation , diminished levels of IFN & #947 ; , TNF & #945 ; , and CCL5 mRNA in pleural effusion cells , and reduced spontaneous lymphocyte proliferation . Together these studies suggest an important role for gpCCL5 in activating leukocytes during M . tuberculosis infection .
URI: http : / /hdl .handle .net /1969 .1 /1506
Date: 2005-02-17

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The role of CCL5 (RANTES) in the immune response against Mycobacterium tuberculosis in the guinea pig. Available electronically from http : / /hdl .handle .net /1969 .1 /1506 .

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