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Differential phenotypic and functional profiles of TcCA-2 -specific cytotoxic CD8+ T cells in the asymptomatic versus cardiac phase in Chagasic patients.

Egui A, Thomas MC, Carrilero B, Segovia M, Alonso C, Marañón C, López MC - PLoS ONE (2015)

Bottom Line: The TcCA-2-specific T CD8+ cells from patients with cardiac symptoms are mainly effector memory cells (TEM and TEMRA) while, those present in the asymptomatic phase are predominantly naive cells (TNAIVE).Moreover, in patients with cardiac symptoms the percentage of cells with senescence features is significantly higher than in patients at the asymptomatic phase of the disease.We consider that the identification of these new class I-restricted epitopes are helpful for designing biomarkers of sickness pathology as well as the development of immunotherapies against T. cruzi infection.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Parasitología y Biomedicina López Neyra, Consejo Superior de Investigaciones Científicas (IPBLN-CSIC), PTS Granada, Avda. del Conocimiento S/N, 18016, Granada, Spain.

ABSTRACT
It has been reported that the immune response mediated by T CD8+ lymphocytes plays a critical role in the control of Trypanosoma cruzi infection and that the clinical symptoms of Chagas disease appear to be related to the competence of the CD8+ T immune response against the parasite. Herewith, in silico prediction and binding assays on TAP-deficient T2 cells were used to identify potential HLA-A*02:01 ligands in the T. cruzi TcCA-2 protein. The TcCA-2-specific CD8+ T cells were functionality evaluated by Granzyme B and cytokine production in peripheral blood mononuclear cells (PBMC) from Chagas disease patients stimulated with the identified HLA-A*02:01 peptides. The specific cells were phenotypically characterized by flow cytometry using several surface markers and HLA-A*02:01 APC-labeled dextramer loaded with the peptides. In the T. cruzi TcCA-2 protein four T CD8+ epitopes were identified which are processed and presented during Chagas disease. Interestingly, a differential cellular phenotypic profile could be correlated with the severity of the disease. The TcCA-2-specific T CD8+ cells from patients with cardiac symptoms are mainly effector memory cells (TEM and TEMRA) while, those present in the asymptomatic phase are predominantly naive cells (TNAIVE). Moreover, in patients with cardiac symptoms the percentage of cells with senescence features is significantly higher than in patients at the asymptomatic phase of the disease. We consider that the identification of these new class I-restricted epitopes are helpful for designing biomarkers of sickness pathology as well as the development of immunotherapies against T. cruzi infection.

No MeSH data available.


Related in: MedlinePlus

Immunophenotyping of memory and differentiation status of the specific CD8+ T cells in patients with Chagas disease.(A) TcCA-2442-451 peptide. (B) TcCA-2607-615 peptide. PBMCs from 6 asymptomatic form patients (IND) and 11 patients with the cardiac form (CCC) were stained for CD45RA, CD27 and CCR7 and analyzed by flow cytometry. According to the combination of antibodies used the CD8 peptide-specific cells were divided in: TNAIVE (CD8+CD45RA+CD27+CCR7+), TEMRA (CD8+CD45RA+CD27-CCR7-), TCM (CD8+CD45RA-CD27+CCR7+), TEM (CD8+CD45RA-CD27-CCR7-), TED (CD8+CD45RA-CD127+), TTD (CD8+CD45RA+CD127-). Median values are represented by horizontal lines. Error bars represent standard deviation intervals. Statistical analyses were carried out using Mann-Whitney U test (#) and Kruskal—Wallis test with Dunn correction (*). Statistically significant differences are indicated (#) or (*) p≤0.05, (# #) or (**) p≤0.01, (# # #) or (***) p≤ 0.001.
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pone.0122115.g003: Immunophenotyping of memory and differentiation status of the specific CD8+ T cells in patients with Chagas disease.(A) TcCA-2442-451 peptide. (B) TcCA-2607-615 peptide. PBMCs from 6 asymptomatic form patients (IND) and 11 patients with the cardiac form (CCC) were stained for CD45RA, CD27 and CCR7 and analyzed by flow cytometry. According to the combination of antibodies used the CD8 peptide-specific cells were divided in: TNAIVE (CD8+CD45RA+CD27+CCR7+), TEMRA (CD8+CD45RA+CD27-CCR7-), TCM (CD8+CD45RA-CD27+CCR7+), TEM (CD8+CD45RA-CD27-CCR7-), TED (CD8+CD45RA-CD127+), TTD (CD8+CD45RA+CD127-). Median values are represented by horizontal lines. Error bars represent standard deviation intervals. Statistical analyses were carried out using Mann-Whitney U test (#) and Kruskal—Wallis test with Dunn correction (*). Statistically significant differences are indicated (#) or (*) p≤0.05, (# #) or (**) p≤0.01, (# # #) or (***) p≤ 0.001.

Mentions: Labeling, using antibodies against CD45RA, CD27 and CCR7 molecules, allowed us to phenotypically characterize TcCA-2442-451 and TcCA-2607-615-specific CD8+ T cells from patients at different phases of the disease (IND and CCC). Thus, as shown in Fig. 3A1 we observed that the TcCA-2442-451-specific CD8+ T cells from IND patients have a higher percentage of cells expressing the TNAIVE phenotype (CD45RA+CD27+CCR7+) than the cells from CCC patients (p≤0.01). However, the percentage of terminal effector memory CD8+ T cells (TEMRA cells, CD45RA+CD27-CCR7-) was significantly lower in IND patients than in CCC (p≤0.05) (Fig. 3A1). Moreover, the TcCA-2442-451-specific CD8+ T cells from CCC patients have a higher percentage of TEMRA cells than that having a phenotype TNAIVE (p≤0.05). Regarding the TcCA-2607-615-specific CD8+ T cells, the percentage of cells expressing CD45RA+CD27+CCR7+ (TNAIVE) was also significantly higher in IND versus CCC patients (p≤0.01) (Fig. 3B1). The percentage of TEM is higher in CCC patients than that in IND patients (p≤0.05). Moreover, the percentage of TEMversus of TCM cells is also higher in CCC patients than that in IND patients (p≤0.05). In fact, the TcCA-2607-615-specific CD8+ T cells from CCC patients had mainly an effector phenotype showing a predominant profile of TEM cells versus of TCM cells (p≤0.05) (Fig. 3B1). By combining the CD45RA and CD127 surface markers it was observed (Fig. 3A2 and 3B2) that both IND and CCC patients have a higher percentage of TcCA-2442-451 and TcCA-2607-615—specific CD8+ T cells at an advanced-stage differentiation (TTD, CD45RA+CD127-) phenotype than at an early-stage differentiation (TED, CD45RA-CD127+) phenotype (p≤0.01).


Differential phenotypic and functional profiles of TcCA-2 -specific cytotoxic CD8+ T cells in the asymptomatic versus cardiac phase in Chagasic patients.

Egui A, Thomas MC, Carrilero B, Segovia M, Alonso C, Marañón C, López MC - PLoS ONE (2015)

Immunophenotyping of memory and differentiation status of the specific CD8+ T cells in patients with Chagas disease.(A) TcCA-2442-451 peptide. (B) TcCA-2607-615 peptide. PBMCs from 6 asymptomatic form patients (IND) and 11 patients with the cardiac form (CCC) were stained for CD45RA, CD27 and CCR7 and analyzed by flow cytometry. According to the combination of antibodies used the CD8 peptide-specific cells were divided in: TNAIVE (CD8+CD45RA+CD27+CCR7+), TEMRA (CD8+CD45RA+CD27-CCR7-), TCM (CD8+CD45RA-CD27+CCR7+), TEM (CD8+CD45RA-CD27-CCR7-), TED (CD8+CD45RA-CD127+), TTD (CD8+CD45RA+CD127-). Median values are represented by horizontal lines. Error bars represent standard deviation intervals. Statistical analyses were carried out using Mann-Whitney U test (#) and Kruskal—Wallis test with Dunn correction (*). Statistically significant differences are indicated (#) or (*) p≤0.05, (# #) or (**) p≤0.01, (# # #) or (***) p≤ 0.001.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4376724&req=5

pone.0122115.g003: Immunophenotyping of memory and differentiation status of the specific CD8+ T cells in patients with Chagas disease.(A) TcCA-2442-451 peptide. (B) TcCA-2607-615 peptide. PBMCs from 6 asymptomatic form patients (IND) and 11 patients with the cardiac form (CCC) were stained for CD45RA, CD27 and CCR7 and analyzed by flow cytometry. According to the combination of antibodies used the CD8 peptide-specific cells were divided in: TNAIVE (CD8+CD45RA+CD27+CCR7+), TEMRA (CD8+CD45RA+CD27-CCR7-), TCM (CD8+CD45RA-CD27+CCR7+), TEM (CD8+CD45RA-CD27-CCR7-), TED (CD8+CD45RA-CD127+), TTD (CD8+CD45RA+CD127-). Median values are represented by horizontal lines. Error bars represent standard deviation intervals. Statistical analyses were carried out using Mann-Whitney U test (#) and Kruskal—Wallis test with Dunn correction (*). Statistically significant differences are indicated (#) or (*) p≤0.05, (# #) or (**) p≤0.01, (# # #) or (***) p≤ 0.001.
Mentions: Labeling, using antibodies against CD45RA, CD27 and CCR7 molecules, allowed us to phenotypically characterize TcCA-2442-451 and TcCA-2607-615-specific CD8+ T cells from patients at different phases of the disease (IND and CCC). Thus, as shown in Fig. 3A1 we observed that the TcCA-2442-451-specific CD8+ T cells from IND patients have a higher percentage of cells expressing the TNAIVE phenotype (CD45RA+CD27+CCR7+) than the cells from CCC patients (p≤0.01). However, the percentage of terminal effector memory CD8+ T cells (TEMRA cells, CD45RA+CD27-CCR7-) was significantly lower in IND patients than in CCC (p≤0.05) (Fig. 3A1). Moreover, the TcCA-2442-451-specific CD8+ T cells from CCC patients have a higher percentage of TEMRA cells than that having a phenotype TNAIVE (p≤0.05). Regarding the TcCA-2607-615-specific CD8+ T cells, the percentage of cells expressing CD45RA+CD27+CCR7+ (TNAIVE) was also significantly higher in IND versus CCC patients (p≤0.01) (Fig. 3B1). The percentage of TEM is higher in CCC patients than that in IND patients (p≤0.05). Moreover, the percentage of TEMversus of TCM cells is also higher in CCC patients than that in IND patients (p≤0.05). In fact, the TcCA-2607-615-specific CD8+ T cells from CCC patients had mainly an effector phenotype showing a predominant profile of TEM cells versus of TCM cells (p≤0.05) (Fig. 3B1). By combining the CD45RA and CD127 surface markers it was observed (Fig. 3A2 and 3B2) that both IND and CCC patients have a higher percentage of TcCA-2442-451 and TcCA-2607-615—specific CD8+ T cells at an advanced-stage differentiation (TTD, CD45RA+CD127-) phenotype than at an early-stage differentiation (TED, CD45RA-CD127+) phenotype (p≤0.01).

Bottom Line: The TcCA-2-specific T CD8+ cells from patients with cardiac symptoms are mainly effector memory cells (TEM and TEMRA) while, those present in the asymptomatic phase are predominantly naive cells (TNAIVE).Moreover, in patients with cardiac symptoms the percentage of cells with senescence features is significantly higher than in patients at the asymptomatic phase of the disease.We consider that the identification of these new class I-restricted epitopes are helpful for designing biomarkers of sickness pathology as well as the development of immunotherapies against T. cruzi infection.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Parasitología y Biomedicina López Neyra, Consejo Superior de Investigaciones Científicas (IPBLN-CSIC), PTS Granada, Avda. del Conocimiento S/N, 18016, Granada, Spain.

ABSTRACT
It has been reported that the immune response mediated by T CD8+ lymphocytes plays a critical role in the control of Trypanosoma cruzi infection and that the clinical symptoms of Chagas disease appear to be related to the competence of the CD8+ T immune response against the parasite. Herewith, in silico prediction and binding assays on TAP-deficient T2 cells were used to identify potential HLA-A*02:01 ligands in the T. cruzi TcCA-2 protein. The TcCA-2-specific CD8+ T cells were functionality evaluated by Granzyme B and cytokine production in peripheral blood mononuclear cells (PBMC) from Chagas disease patients stimulated with the identified HLA-A*02:01 peptides. The specific cells were phenotypically characterized by flow cytometry using several surface markers and HLA-A*02:01 APC-labeled dextramer loaded with the peptides. In the T. cruzi TcCA-2 protein four T CD8+ epitopes were identified which are processed and presented during Chagas disease. Interestingly, a differential cellular phenotypic profile could be correlated with the severity of the disease. The TcCA-2-specific T CD8+ cells from patients with cardiac symptoms are mainly effector memory cells (TEM and TEMRA) while, those present in the asymptomatic phase are predominantly naive cells (TNAIVE). Moreover, in patients with cardiac symptoms the percentage of cells with senescence features is significantly higher than in patients at the asymptomatic phase of the disease. We consider that the identification of these new class I-restricted epitopes are helpful for designing biomarkers of sickness pathology as well as the development of immunotherapies against T. cruzi infection.

No MeSH data available.


Related in: MedlinePlus