Severity: Warning
Message: file_get_contents(https://...@pubfacts.com&api_key=b8daa3ad693db53b1410957c26c9a51b4908&a=1): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests
Filename: helpers/my_audit_helper.php
Line Number: 176
Backtrace:
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 176
Function: file_get_contents
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 250
Function: simplexml_load_file_from_url
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 1034
Function: getPubMedXML
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 3152
Function: GetPubMedArticleOutput_2016
File: /var/www/html/application/controllers/Detail.php
Line: 575
Function: pubMedSearch_Global
File: /var/www/html/application/controllers/Detail.php
Line: 489
Function: pubMedGetRelatedKeyword
File: /var/www/html/index.php
Line: 316
Function: require_once
Aim: To explore whether the human peripheral gammadelta T cells stimulated by polypeptide heat resistant antigen from Mycobacterium tuberculosis (Mtb-HAg) express phenotype and have the fuction of antigen presentation.
Methods: The monocytes isolated by adhesion method from peripheral blood mononuclear cells (PBMCs) were cultured with GM-CSF, IL-4 and induced into the mature dendritic cells by adding LPS. PBMCs were stimulated with Mtb-HAg and IL-2 to expand predominantally gammadelta T cells that were further purified by flow sorting. The pure T cells were isolated from PBMCs using adhesion revomal and nylon column method, labeled with CFSE, and cultured in alone with Mtb secretory antigen (Mtb-SAg), in monocytes with Mtb-SAg, or with Mtb-SAg pre-pulsed dendritic cells or Mtb-HAg activated gammadelta T cells for 11 days. The proliferation of the CD4(+) T cells were measured by flowcytometry. The Mtb-HAg activated gammadelta T cells were incubated with FITC labelled Mtb-SAg for different time, and the ingestion of Mtb-SAg by gammadelta T cells was measured by flowcytometry and observed with laser confocal microscopye.
Results: The expressions of MHC-II and co-stimulatory molecules CD80 and CD86 on gammadelta T cells that activated by Mtb-HAg marketly incrased in comparison with that on the resting gammadelta T cells. The expanded cell counts and proliferation index of the pure naïve T cells that induced by Mtb-SAg pretreated activated gammadelta T cells were similar to that induced by mature DC. By using flowcytometer and laser confocal microscope, FITC labellaed Mtb-SAg was ingested by the activated gammadelta T cells.
Conclusion: gammadelta T cells stimulated by polypeptide Mtb-HAg express the phenotype of professional antigen-presenting cells and they are able to present Mtb-SAg to naïve T cells to induce the proliferation of CD4(+) T cells.
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