The kinetics of expansion and contraction of the double positive CxCR5+Bcl-6+ CD4 T cell population closely paralleled the development and resolution of germinal centers in the spleen. The relative transcript LPA2 antagonist 1 levels from total SMCs (upper panels) and total lymph node cells (lower panels) were determined by qPCR in non-infected animals and after 11, 28 and 250 days of contamination. Results are shown as mean SEM of the fold change over the noninfected samples, which were attributed a normalized value of 1 1. (A) and (I) and in sorted lymph node CD4 T cells were determined by qPCR. Results are shown as fold change SEM over non-infected samples. (B) Representative density plots depicting the expression of CXCR5 and Bcl-6 (upper panels) or CXCR5 and PD-1 (lower panels) in lymph node CD4 T cells during the course of contamination. (C) Expression (mean RGS1 SEM) of CXCR5, Bcl-6 and PD-1 among splenic CD4 T cells during the course of contamination. (D) Percentage (mean SEM) of expression of the double positive CXCR5+Bcl-6+ or CXCR5+PD-1+ populations and the triple positive CXCR5+Bcl-6+PD-1+ populace among lymph node CD4 T cells. Statistical analysis was performed by one-way ANOVA, followed by a Bonferroni’s post-hoc test.(TIF) ppat.1004096.s008.tif (1.7M) GUID:?1C687998-BAC8-47E9-85CF-F0CD66B38B19 Figure S9: Follicular helper T cell imaging in LPA2 antagonist 1 lymph nodes during infection of rhesus macaques. (ACD) Lymph node tissue sections were stained with antibodies against CXCR5 (blue), CD4 (green) and PD-1 (red) and imaged by confocal microscopy. Shown are representative pictures of a na?ve animal LPA2 antagonist 1 (A) and at 11 (B), 28 (C) and 250 (D) days after infection. (E) Inset from physique S8D as defined by the white square.(TIF) ppat.1004096.s009.tif (9.2M) GUID:?614FE6EC-B1C1-4BE1-A53A-1B3FABFF2570 Figure S10: QPCR products were separated in a 2% agarose gel. The 100 bp DNA markers are shown alongside the bands.(TIF) ppat.1004096.s010.tif (2.0M) GUID:?C2A69454-61F1-42AD-BD25-6E0D2F057931 Table S1: Information related to the antibodies used in flow cytometry and tissue immunofluorescence studies.(DOCX) ppat.1004096.s011.docx (14K) GUID:?1FF02856-94F6-48B5-8595-EDCF17425705 Table S2: Sequence, PCR product size and accession number of the primers used in this study.(DOCX) ppat.1004096.s012.docx (16K) GUID:?A31DEDB2-B72A-476A-9874-CE2355B09112 Material and Methods S1: Detailed description of the protocols employed for quantification of serum analytes.(DOCX) ppat.1004096.s013.docx (16K) GUID:?AF27702D-E3A7-430C-87A6-EFA6F67DBC06 Abstract causes a chronic infectious disease named visceral leishmaniasis (VL). We employed a non-human primate model to monitor immune parameters over time and gain new insights into the disease. Rhesus macaques were infected with and the T helper and B cell immunological profiles characterized during acute and chronic phases of contamination. Parasite detection in visceral compartments during the acute phase was associated with differentiation of effector memory CD4 T cells and increased levels of Th1 transcripts. At the chronic phase, parasites colonized novel lymphoid niches concomitant with increased expression of promastigotes in rhesus macaques and followed the animals for a period of eight months. In this model, parasites dock to the liver and spleen shortly after inoculation and remain in these visceral compartments during all the acute phase of contamination. However, at the chronic phase, additional body locations appeared colonized (lymph nodes, bone marrow). During the acute phase, a Th1-polarized CD4 T cell response develops in the spleen, but, and concomitant with parasite growth, it waned at the chronic phase. Furthermore, we observed the acute expansion of a splenic T follicular helper (Tfh) cell populace, a CD4+ T cell subset specialized to assist B cells in the production of antigen-specific antibody. These cells were localized in close association with B cell follicles but, interestingly, the Tfh populace is lost at the chronic phase. Nevertheless, there was a close association between the development of Tfh cells and the differentiation of B cells that produce or species and develop a life-long latent contamination [5], contrasting with the potentially fatal human VL in which progressive illness develops, even in the presence of detectable levels of IFN- and TNF.
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