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Peripheral myeloid-derived suppressor and T regulatory PD-1 positive cells predict response to neoadjuvant short-course radiotherapy in rectal cancer patients.

Napolitano M, D'Alterio C, Cardone E, Trotta AM, Pecori B, Rega D, Pace U, Scala D, Scognamiglio G, Tatangelo F, Cacciapuoti C, Pacelli R, Delrio P, Scala S - Oncotarget (2015)

Bottom Line: Peripheral Granulocytic-MDSCs (G-MDSC) [LIN-/HLA-DR-/CD11b+/CD14-/CD15+/CD33+], Monocytic (M-MDSC) [CD14+/HLA-DR-/lowCD11b+/CD33+] and Tregs [CD4+/CD25hi+/FOXP3+- CTLA-4/PD1] basal value was significantly higher in LARC patients compared to healthy donors (HD).In this pilot study MDSCs and Tregs decrease during the SC-RT treatment could represent a biomarker of response in LARC patients.Further studies are needed to confirm that the deepest M-MDSC reduction and increase in Treg-PD1 cells within 5-8 weeks from the beginning of treatment could discriminate LARC patients poor responding to SC-RT.

View Article: PubMed Central - PubMed

Affiliation: Immunology Unit, Istituto Nazionale per lo Studio e la Cura dei Tumori "Fondazione Giovanni Pascale" - I.R.C.C.S., Naples, Italy.

ABSTRACT
Short-course preoperative radiotherapy (SC-RT) followed by total mesorectal excision (TME) is one therapeutic option for locally advanced rectal cancer (LARC) patients. Since radio-induced DNA damage may affect tumor immunogenicity, Myeloid-derived suppressor cells (MDSCs) and T regulatory cells (Tregs) were evaluated in 13 patients undergoing SC-RT and TME for LARC. Peripheral Granulocytic-MDSCs (G-MDSC) [LIN-/HLA-DR-/CD11b+/CD14-/CD15+/CD33+], Monocytic (M-MDSC) [CD14+/HLA-DR-/lowCD11b+/CD33+] and Tregs [CD4+/CD25hi+/FOXP3+- CTLA-4/PD1] basal value was significantly higher in LARC patients compared to healthy donors (HD). Peripheral MDSC and Tregs were evaluated at time 0 (T0), after 2 and 5 weeks (T2-T5) from radiotherapy; before surgery (T8) and 6-12 months after surgery (T9, T10). G-MDSC decreased at T5 and further at T8 while M-MDSC cells decreased at T5; Tregs reached the lowest value at T5. LARC poor responder patients displayed a major decrease in M-MDSC after SC-RT and an increase of Treg-PD-1. In this pilot study MDSCs and Tregs decrease during the SC-RT treatment could represent a biomarker of response in LARC patients. Further studies are needed to confirm that the deepest M-MDSC reduction and increase in Treg-PD1 cells within 5-8 weeks from the beginning of treatment could discriminate LARC patients poor responding to SC-RT.

No MeSH data available.


Related in: MedlinePlus

Flow cytometry analysis of Granulocytic (G-MDSc LIN-/HLA-DR−/CD11b+/CD14−/CD15+/CD33+) and Monocytic (M-MDSc CD14+/HLADR−/low/CD11b+CD33+)Peripheral blood from healthy donors (A, B) and LARC patients (C, D) were stained for the G-MDSC markers Lineage 1 (CD3/CD14/CD19/CD20/CD56), HLA-DR, CD33, CD11b, and CD15 (A, C) and the M-MDSC markers CD14 and HLA-DR (B, D). Representative dot plot is shown. Gates were set based on isotype controls. Numbers represent the percentages from the parental populations gated. The gating strategy used to analyze the samples is illustrated. Acquired cells were first gated (Pe-Cy7/Fitc subset) based on the expression of LIN1 and HLA-DR. Within this population the fraction of cells G-MDSc expressing both CD11b, CD14, CD33 and CD15 was determined. Therefore, G-MDSC were defined as LIN−/HLA-DR−/low/CD11b+/CD14−/CD15+/CD33+ cells (A and C). Acquired cells were first gated (Pe/Fitc subset) based on the expression of LIN1 and CD11b. Within this population the fraction of cells M-MDSc expressing both CD14, CD15, HLA-DR, and CD33 was determined. Therefore, M-MDSC was defined as CD14+/HLA-DR−/low/CD11b+/CD33+cells (B and C). MDSCs percentage was calculated as percentage of total nucleated cells in whole blood samples.
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Figure 2: Flow cytometry analysis of Granulocytic (G-MDSc LIN-/HLA-DR−/CD11b+/CD14−/CD15+/CD33+) and Monocytic (M-MDSc CD14+/HLADR−/low/CD11b+CD33+)Peripheral blood from healthy donors (A, B) and LARC patients (C, D) were stained for the G-MDSC markers Lineage 1 (CD3/CD14/CD19/CD20/CD56), HLA-DR, CD33, CD11b, and CD15 (A, C) and the M-MDSC markers CD14 and HLA-DR (B, D). Representative dot plot is shown. Gates were set based on isotype controls. Numbers represent the percentages from the parental populations gated. The gating strategy used to analyze the samples is illustrated. Acquired cells were first gated (Pe-Cy7/Fitc subset) based on the expression of LIN1 and HLA-DR. Within this population the fraction of cells G-MDSc expressing both CD11b, CD14, CD33 and CD15 was determined. Therefore, G-MDSC were defined as LIN−/HLA-DR−/low/CD11b+/CD14−/CD15+/CD33+ cells (A and C). Acquired cells were first gated (Pe/Fitc subset) based on the expression of LIN1 and CD11b. Within this population the fraction of cells M-MDSc expressing both CD14, CD15, HLA-DR, and CD33 was determined. Therefore, M-MDSC was defined as CD14+/HLA-DR−/low/CD11b+/CD33+cells (B and C). MDSCs percentage was calculated as percentage of total nucleated cells in whole blood samples.

Mentions: Previous studies have described significant increase in circulating Lin−/HLADR−/CD11b+CD33+ MDSCs in the peripheral blood of patients with advanced cancer including colon cancer [21]. With the intent to identify peripheral pattern of immune modulation in response to treatment, a pilot study was conducted on 13 LARC patients subjected to neo adjuvant SC-RT (5 fractions of 5Gy for 5 days) and then surgery (Table 1). MDSC and Treg cells were evaluated at time 0 (T0), after two weeks from the beginning of RT (T2), after 5 weeks from the beginning of radiotherapy (T5); before surgery (T8),6 (T9) and 12 (T10) months after surgery (Figure 1). In Figure 2 it is shown a representative flow cytometry analysis on peripheral blood from healthy individual (Figure 2A–2B) and a LARC patient (Figure 2C–2D) for G-MDSC (LIN−/HLA-DR−/CD11b+/CD14−/CD15+/CD33+and M-MDSC (CD14+/HLA-DR−/low/CD11b+/CD33+). As previously reported [21], a significantly higher number of circulating G-MDSCs (mean 1,85% vs 0,56%; p = 0.0026) and M-MDSC (mean 0,66% vs 0,31%; p = 0.0106) was detected in LARC patients at time 0 compared to healthy donors (Figure 3A–3B); also Tregs (CD4+/CD25hi+/FoxP3+/CTLA4+ and CD4+/CD25hi+/FoxP3+/PD1+) were higher in LARC patients compared to healthy donors (mean 0.18% vs 0.29% and mean 0.03% vs. 0.14%; p = 0.0340) (Figure 3C–3D). The MDSCs and Tregs course over treatment was then evaluated in 13 patients undergone to SC-RT followed by surgery. G-MDSC decreased at T5 and further at T8 while M-MDSC cells decreased at T5 being stable up to T8 (Figure 4A–4B); Tregs reached the lowest value at five weeks after the beginning of SC-RT (Figure 4C–4D). Tregs variations are not related to CD4 total population changes at the indicated time points (Supplementary Figure 1).


Peripheral myeloid-derived suppressor and T regulatory PD-1 positive cells predict response to neoadjuvant short-course radiotherapy in rectal cancer patients.

Napolitano M, D'Alterio C, Cardone E, Trotta AM, Pecori B, Rega D, Pace U, Scala D, Scognamiglio G, Tatangelo F, Cacciapuoti C, Pacelli R, Delrio P, Scala S - Oncotarget (2015)

Flow cytometry analysis of Granulocytic (G-MDSc LIN-/HLA-DR−/CD11b+/CD14−/CD15+/CD33+) and Monocytic (M-MDSc CD14+/HLADR−/low/CD11b+CD33+)Peripheral blood from healthy donors (A, B) and LARC patients (C, D) were stained for the G-MDSC markers Lineage 1 (CD3/CD14/CD19/CD20/CD56), HLA-DR, CD33, CD11b, and CD15 (A, C) and the M-MDSC markers CD14 and HLA-DR (B, D). Representative dot plot is shown. Gates were set based on isotype controls. Numbers represent the percentages from the parental populations gated. The gating strategy used to analyze the samples is illustrated. Acquired cells were first gated (Pe-Cy7/Fitc subset) based on the expression of LIN1 and HLA-DR. Within this population the fraction of cells G-MDSc expressing both CD11b, CD14, CD33 and CD15 was determined. Therefore, G-MDSC were defined as LIN−/HLA-DR−/low/CD11b+/CD14−/CD15+/CD33+ cells (A and C). Acquired cells were first gated (Pe/Fitc subset) based on the expression of LIN1 and CD11b. Within this population the fraction of cells M-MDSc expressing both CD14, CD15, HLA-DR, and CD33 was determined. Therefore, M-MDSC was defined as CD14+/HLA-DR−/low/CD11b+/CD33+cells (B and C). MDSCs percentage was calculated as percentage of total nucleated cells in whole blood samples.
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Related In: Results  -  Collection

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Figure 2: Flow cytometry analysis of Granulocytic (G-MDSc LIN-/HLA-DR−/CD11b+/CD14−/CD15+/CD33+) and Monocytic (M-MDSc CD14+/HLADR−/low/CD11b+CD33+)Peripheral blood from healthy donors (A, B) and LARC patients (C, D) were stained for the G-MDSC markers Lineage 1 (CD3/CD14/CD19/CD20/CD56), HLA-DR, CD33, CD11b, and CD15 (A, C) and the M-MDSC markers CD14 and HLA-DR (B, D). Representative dot plot is shown. Gates were set based on isotype controls. Numbers represent the percentages from the parental populations gated. The gating strategy used to analyze the samples is illustrated. Acquired cells were first gated (Pe-Cy7/Fitc subset) based on the expression of LIN1 and HLA-DR. Within this population the fraction of cells G-MDSc expressing both CD11b, CD14, CD33 and CD15 was determined. Therefore, G-MDSC were defined as LIN−/HLA-DR−/low/CD11b+/CD14−/CD15+/CD33+ cells (A and C). Acquired cells were first gated (Pe/Fitc subset) based on the expression of LIN1 and CD11b. Within this population the fraction of cells M-MDSc expressing both CD14, CD15, HLA-DR, and CD33 was determined. Therefore, M-MDSC was defined as CD14+/HLA-DR−/low/CD11b+/CD33+cells (B and C). MDSCs percentage was calculated as percentage of total nucleated cells in whole blood samples.
Mentions: Previous studies have described significant increase in circulating Lin−/HLADR−/CD11b+CD33+ MDSCs in the peripheral blood of patients with advanced cancer including colon cancer [21]. With the intent to identify peripheral pattern of immune modulation in response to treatment, a pilot study was conducted on 13 LARC patients subjected to neo adjuvant SC-RT (5 fractions of 5Gy for 5 days) and then surgery (Table 1). MDSC and Treg cells were evaluated at time 0 (T0), after two weeks from the beginning of RT (T2), after 5 weeks from the beginning of radiotherapy (T5); before surgery (T8),6 (T9) and 12 (T10) months after surgery (Figure 1). In Figure 2 it is shown a representative flow cytometry analysis on peripheral blood from healthy individual (Figure 2A–2B) and a LARC patient (Figure 2C–2D) for G-MDSC (LIN−/HLA-DR−/CD11b+/CD14−/CD15+/CD33+and M-MDSC (CD14+/HLA-DR−/low/CD11b+/CD33+). As previously reported [21], a significantly higher number of circulating G-MDSCs (mean 1,85% vs 0,56%; p = 0.0026) and M-MDSC (mean 0,66% vs 0,31%; p = 0.0106) was detected in LARC patients at time 0 compared to healthy donors (Figure 3A–3B); also Tregs (CD4+/CD25hi+/FoxP3+/CTLA4+ and CD4+/CD25hi+/FoxP3+/PD1+) were higher in LARC patients compared to healthy donors (mean 0.18% vs 0.29% and mean 0.03% vs. 0.14%; p = 0.0340) (Figure 3C–3D). The MDSCs and Tregs course over treatment was then evaluated in 13 patients undergone to SC-RT followed by surgery. G-MDSC decreased at T5 and further at T8 while M-MDSC cells decreased at T5 being stable up to T8 (Figure 4A–4B); Tregs reached the lowest value at five weeks after the beginning of SC-RT (Figure 4C–4D). Tregs variations are not related to CD4 total population changes at the indicated time points (Supplementary Figure 1).

Bottom Line: Peripheral Granulocytic-MDSCs (G-MDSC) [LIN-/HLA-DR-/CD11b+/CD14-/CD15+/CD33+], Monocytic (M-MDSC) [CD14+/HLA-DR-/lowCD11b+/CD33+] and Tregs [CD4+/CD25hi+/FOXP3+- CTLA-4/PD1] basal value was significantly higher in LARC patients compared to healthy donors (HD).In this pilot study MDSCs and Tregs decrease during the SC-RT treatment could represent a biomarker of response in LARC patients.Further studies are needed to confirm that the deepest M-MDSC reduction and increase in Treg-PD1 cells within 5-8 weeks from the beginning of treatment could discriminate LARC patients poor responding to SC-RT.

View Article: PubMed Central - PubMed

Affiliation: Immunology Unit, Istituto Nazionale per lo Studio e la Cura dei Tumori "Fondazione Giovanni Pascale" - I.R.C.C.S., Naples, Italy.

ABSTRACT
Short-course preoperative radiotherapy (SC-RT) followed by total mesorectal excision (TME) is one therapeutic option for locally advanced rectal cancer (LARC) patients. Since radio-induced DNA damage may affect tumor immunogenicity, Myeloid-derived suppressor cells (MDSCs) and T regulatory cells (Tregs) were evaluated in 13 patients undergoing SC-RT and TME for LARC. Peripheral Granulocytic-MDSCs (G-MDSC) [LIN-/HLA-DR-/CD11b+/CD14-/CD15+/CD33+], Monocytic (M-MDSC) [CD14+/HLA-DR-/lowCD11b+/CD33+] and Tregs [CD4+/CD25hi+/FOXP3+- CTLA-4/PD1] basal value was significantly higher in LARC patients compared to healthy donors (HD). Peripheral MDSC and Tregs were evaluated at time 0 (T0), after 2 and 5 weeks (T2-T5) from radiotherapy; before surgery (T8) and 6-12 months after surgery (T9, T10). G-MDSC decreased at T5 and further at T8 while M-MDSC cells decreased at T5; Tregs reached the lowest value at T5. LARC poor responder patients displayed a major decrease in M-MDSC after SC-RT and an increase of Treg-PD-1. In this pilot study MDSCs and Tregs decrease during the SC-RT treatment could represent a biomarker of response in LARC patients. Further studies are needed to confirm that the deepest M-MDSC reduction and increase in Treg-PD1 cells within 5-8 weeks from the beginning of treatment could discriminate LARC patients poor responding to SC-RT.

No MeSH data available.


Related in: MedlinePlus