J Gynecol Oncol.  2010 Mar;21(1):38-44. 10.3802/jgo.2010.21.1.38.

Proliferation of CD4(+)CD25(high+)Foxp3(+) regulatory T lymphocytes in ex vivo expanded ascitic fluid from primary and recurrent ovarian carcinoma

• Lee SW
• Kim YM
• Lee HY
• Kim DY
• Kim JH
• Nam JH
• Kim YT

• KMID: 2173510
• DOI: http://doi.org/10.3802/jgo.2010.21.1.38

Abstract

OBJECTIVE
Regulatory T lymphocytes evoke the immune tolerance by suppressing and inactivating cytotoxic T lymphocytes. The objective of this study was to compare the proportion of regulatory T lymphocytes, precisely defined as CD4(+)CD25(high+)Foxp3(+) T lymphocytes, in primary and recurrent ovarian carcinoma before and after ex vivo expansion of ascites with interleukin-2 (IL-2). METHODS: Ascitic fluid samples were obtained from 26 patients with ovarian carcinoma. Lymphocytes were isolated from ascites and cell markers were analyzed by flow cytometry using anti-CD3/CD4/CD8/CD16/CD56/CD25 and anti-Foxp3 antibodies. Lymphocytes were incubated for 2 to 3 weeks and expanded ex vivo by IL-2 stimulation and their phenotypes were analyzed by flow cytometry. RESULTS: Following ex vivo expansion, ascitic fluid lymphocytes increased by a greater extent in the recurrent group than in the primary group. The proportion of ex vivo-expanded lymphocytes changed as follows; CD4(+) T lymphocytes increased, CD8(+) T lymphocytes decreased, and the proportion of CD3(-)CD16(+)56(+) NK cells was unchanged. The proportion of CD4(+)CD25(high+)Foxp3(+) regulatory T lymphocytes in CD4(+) T lymphocytes increased after ex vivo expansion in both groups, but to a greater degree in the recurrent group. CONCLUSION: This study showed that regulatory T lymphocytes, neither cytotoxic T lymphocytes nor NK cells, were extensively increased after ex vivo expansion, especially in recurrent ovarian carcinoma. These results may provide information that helps to guide the future development of adoptive immunotherapy against ovarian carcinoma.

Keyword

Regulatory T lymphocyte; Foxp3; Ex vivo expansion; Ovarian carcinoma; Ascites

MeSH Terms

Antibodies
Ascites
Ascitic Fluid
Flow Cytometry
Humans
Immune Tolerance
Immunotherapy, Adoptive
Interleukin-2
Killer Cells, Natural
Lymphocytes
Phenotype
T-Lymphocytes
T-Lymphocytes, Cytotoxic
T-Lymphocytes, Regulatory
Antibodies
Interleukin-2

Figure

• 

  Fig. 1 The proportion of each immune cells before and after ex vivo expansion. CD4+ T lymphocytes were increased and CD8+ T lymphocytes were decreased after ex vivo expansion. The change in NK cells was not significant statistically. CD4+CD25high+Foxp3+ regulatory T lymphocytes (last pair of bars) increased significantly after ex vivo expansion.

• 

  Fig. 2 The proportional change of immune cells in primary and recurrent ovarian carcinoma after ex vivo expansion. CD4+ T lymphocytes, CD8+ T lymphocytes and NK cells did not differ significantly between primary and recurrent ovarian carcinoma. CD4+CD25high+Foxp3+ regulatory T lymphocytes was significantly greater in recurrent ovarian carcinoma than in primary ovarian carcinoma.

• 

  Fig. 3 Flow cytometry analysis of lymphocytes in primary and recurrent ovarian carcinoma after ex vivo expansion. (A) Representative primary ovarian carcinoma. (B) Representative recurrent ovarian carcinoma.

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