2003
DOI: 10.1016/s0022-1759(03)00255-2
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Optimization of methods to assess human mucosal T-cell responses to HIV infection

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Cited by 94 publications
(110 citation statements)
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“…Lamina propria lymphocytes (LPLs) were isolated from antral biopsy specimens by collagenase type II (Sigma) digestion (16). Samples were incubated in 10 ml RPMI 1640 (Life Technologies by Life Technologies) containing 7.5% FCS (Sigma), 0.5 mg/ml collagenase type II-S (sterile filtered) (clostridiopeptidase A derived from Clostridium histolyticum; Sigma), 0.1 mg/ml trypsin inhibitor (from Glycine max; Sigma), and 0.1 mg/ml DNase I (Roche, Berlin, Germany) for 15 min at 37˚C and 5% CO 2 , with intermittent shaking.…”
Section: Preparation Of Lamina Propria Lymphocytesmentioning
confidence: 99%
“…Lamina propria lymphocytes (LPLs) were isolated from antral biopsy specimens by collagenase type II (Sigma) digestion (16). Samples were incubated in 10 ml RPMI 1640 (Life Technologies by Life Technologies) containing 7.5% FCS (Sigma), 0.5 mg/ml collagenase type II-S (sterile filtered) (clostridiopeptidase A derived from Clostridium histolyticum; Sigma), 0.1 mg/ml trypsin inhibitor (from Glycine max; Sigma), and 0.1 mg/ml DNase I (Roche, Berlin, Germany) for 15 min at 37˚C and 5% CO 2 , with intermittent shaking.…”
Section: Preparation Of Lamina Propria Lymphocytesmentioning
confidence: 99%
“…Although data on mucosal HIV-specific immunity during the acute phase of infection are scant, recent studies have investigated HIV-specific T-cell immunity in rectal biopsies of chronically HIV-infected individuals. 63,64 Initially, these studies were restricted to the physical examination of HIVspecific CD8 T cells using peptide-major histocompatibility complex tetramer technology. 63 These studies demonstrated that HIV-specific CD8 T cells were detectable in the GI tract and that the frequency of HIV-specific CD8 T cells in the GI tract was similar to that observed in peripheral blood, approximately 1% of CD8 T cells.…”
Section: Gastrointestinal Hiv-specific Immune Responsesmentioning
confidence: 99%
“…63,64 Initially, these studies were restricted to the physical examination of HIVspecific CD8 T cells using peptide-major histocompatibility complex tetramer technology. 63 These studies demonstrated that HIV-specific CD8 T cells were detectable in the GI tract and that the frequency of HIV-specific CD8 T cells in the GI tract was similar to that observed in peripheral blood, approximately 1% of CD8 T cells. 40 However, this is an overestimate, since these studies did not express the frequency of memory CD8 T cells, which are HIV-specific; moreover, almost all the GI-tract CD8 T cells belong to the memory compartment, yet naive CD8 T cells circulate in peripheral blood.…”
Section: Gastrointestinal Hiv-specific Immune Responsesmentioning
confidence: 99%
“…A suspension of mucosal cells from rectosigmoid biopsies was obtained after 2 successive 0.25-mg/mL collagenase type II treatments (Sigma-Aldrich), as previously described. 22 …”
Section: Tissue Collection and Processingmentioning
confidence: 99%