Limits...
FOXP3 and CTLA4 overexpression in multiple myeloma bone marrow as a sign of accumulation of CD4(+) T regulatory cells.

Braga WM, da Silva BR, de Carvalho AC, Maekawa YH, Bortoluzzo AB, Rizzatti EG, Atanackovic D, Colleoni GW - Cancer Immunol. Immunother. (2014)

Bottom Line: RORγt expression was similar in MM patients and controls.Overexpression of FOXP3 and CTLA4 in total BM samples suggests a local accumulation of immunosuppressive Tregs, the MM tumor environment, possibly dampening anti-tumor host immune responses.Therapeutic approaches targeting Treg cells and restoring local anti-tumor immunity may provide new treatment strategies for this incurable malignancy.

View Article: PubMed Central - PubMed

Affiliation: Universidade Federal de São Paulo [UNIFESP], Rua Diogo de Faria, 824, 5º andar, Hemocentro, São Paulo, CEP 04037-003, Brazil, wmoises@uol.com.br.

ABSTRACT

Introduction: Multiple myeloma (MM) development involves a series of genetic abnormalities and changes in the bone marrow (BM) microenvironment, favoring the growth of the tumor and failure of local immune control. T regulatory (Treg) cells play an important role in dampening anti-tumor immune responses while T-helper-17 (Th17) cells seem to be critical for the eradication of malignant cells. The aim of our study was to characterize the expression of Treg- and Th17-related genes in total myeloma BM samples to assess their role as biomarkers, prognostic factors, and possible therapeutic targets in this incurable disease.

Methods: Expression of markers for Treg (FOXP3, CTLA4) and Th17 cells (RORγt) was determined by quantitative real-time PCR in BM aspirates of 46 MM patients, four patients with monoclonal gammopathy of undetermined significance, five solitary plasmacytomas, and five healthy BM donors. Gene expression was evaluated regarding an influence on the patients' overall survival (OS).

Results: FOXP3 and CTLA4 presented a sixfold (p = 0.02) and 30-fold higher expression (p = 0.03), respectively, in MM patients than in controls. RORγt expression was similar in MM patients and controls. Median OS of MM patients was 16.8 (range 4.5-29.1) months, and international staging system was the only independent prognostic factor for patients survival.

Conclusions: Overexpression of FOXP3 and CTLA4 in total BM samples suggests a local accumulation of immunosuppressive Tregs, the MM tumor environment, possibly dampening anti-tumor host immune responses. Therapeutic approaches targeting Treg cells and restoring local anti-tumor immunity may provide new treatment strategies for this incurable malignancy.

Show MeSH

Related in: MedlinePlus

Multiparameter flow cytometry analysis showing CD4+ CD25high immunophenotype in bone marrow aspirate samples from a case of multiple myeloma (a) and a healthy donor (c) (indicated as color dots); FOXP3+CTLA4(CD152) positive (color dots) in the same myeloma sample (b) and negative in the same healthy donor sample (D)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4209089&req=5

Fig2: Multiparameter flow cytometry analysis showing CD4+ CD25high immunophenotype in bone marrow aspirate samples from a case of multiple myeloma (a) and a healthy donor (c) (indicated as color dots); FOXP3+CTLA4(CD152) positive (color dots) in the same myeloma sample (b) and negative in the same healthy donor sample (D)

Mentions: Next, we analyzed whether real-time PCR would be as useful for quantification of immunomodulating T cell subsets as flow cytometry (considered the gold standard method in most of the previous studies). Applying both methods to 19 BM samples (13 cases of MM, 1 SP, and 2 MGUS patients), as well as 3 healthy donors, we found a linear correlation (r = 0.48; p = 0.03) between the proportion of Tregs (CD3+CD4+CD25highFOXP3+CTLA4+) in total BM samples as determined by flow cytometry and the expression of CTLA4 assessed by qPCR (Figs. 2, 3a and Supplementary Figure 1).Fig. 2


FOXP3 and CTLA4 overexpression in multiple myeloma bone marrow as a sign of accumulation of CD4(+) T regulatory cells.

Braga WM, da Silva BR, de Carvalho AC, Maekawa YH, Bortoluzzo AB, Rizzatti EG, Atanackovic D, Colleoni GW - Cancer Immunol. Immunother. (2014)

Multiparameter flow cytometry analysis showing CD4+ CD25high immunophenotype in bone marrow aspirate samples from a case of multiple myeloma (a) and a healthy donor (c) (indicated as color dots); FOXP3+CTLA4(CD152) positive (color dots) in the same myeloma sample (b) and negative in the same healthy donor sample (D)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4209089&req=5

Fig2: Multiparameter flow cytometry analysis showing CD4+ CD25high immunophenotype in bone marrow aspirate samples from a case of multiple myeloma (a) and a healthy donor (c) (indicated as color dots); FOXP3+CTLA4(CD152) positive (color dots) in the same myeloma sample (b) and negative in the same healthy donor sample (D)
Mentions: Next, we analyzed whether real-time PCR would be as useful for quantification of immunomodulating T cell subsets as flow cytometry (considered the gold standard method in most of the previous studies). Applying both methods to 19 BM samples (13 cases of MM, 1 SP, and 2 MGUS patients), as well as 3 healthy donors, we found a linear correlation (r = 0.48; p = 0.03) between the proportion of Tregs (CD3+CD4+CD25highFOXP3+CTLA4+) in total BM samples as determined by flow cytometry and the expression of CTLA4 assessed by qPCR (Figs. 2, 3a and Supplementary Figure 1).Fig. 2

Bottom Line: RORγt expression was similar in MM patients and controls.Overexpression of FOXP3 and CTLA4 in total BM samples suggests a local accumulation of immunosuppressive Tregs, the MM tumor environment, possibly dampening anti-tumor host immune responses.Therapeutic approaches targeting Treg cells and restoring local anti-tumor immunity may provide new treatment strategies for this incurable malignancy.

View Article: PubMed Central - PubMed

Affiliation: Universidade Federal de São Paulo [UNIFESP], Rua Diogo de Faria, 824, 5º andar, Hemocentro, São Paulo, CEP 04037-003, Brazil, wmoises@uol.com.br.

ABSTRACT

Introduction: Multiple myeloma (MM) development involves a series of genetic abnormalities and changes in the bone marrow (BM) microenvironment, favoring the growth of the tumor and failure of local immune control. T regulatory (Treg) cells play an important role in dampening anti-tumor immune responses while T-helper-17 (Th17) cells seem to be critical for the eradication of malignant cells. The aim of our study was to characterize the expression of Treg- and Th17-related genes in total myeloma BM samples to assess their role as biomarkers, prognostic factors, and possible therapeutic targets in this incurable disease.

Methods: Expression of markers for Treg (FOXP3, CTLA4) and Th17 cells (RORγt) was determined by quantitative real-time PCR in BM aspirates of 46 MM patients, four patients with monoclonal gammopathy of undetermined significance, five solitary plasmacytomas, and five healthy BM donors. Gene expression was evaluated regarding an influence on the patients' overall survival (OS).

Results: FOXP3 and CTLA4 presented a sixfold (p = 0.02) and 30-fold higher expression (p = 0.03), respectively, in MM patients than in controls. RORγt expression was similar in MM patients and controls. Median OS of MM patients was 16.8 (range 4.5-29.1) months, and international staging system was the only independent prognostic factor for patients survival.

Conclusions: Overexpression of FOXP3 and CTLA4 in total BM samples suggests a local accumulation of immunosuppressive Tregs, the MM tumor environment, possibly dampening anti-tumor host immune responses. Therapeutic approaches targeting Treg cells and restoring local anti-tumor immunity may provide new treatment strategies for this incurable malignancy.

Show MeSH
Related in: MedlinePlus