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WAM Essentials, Inc.
Systemic Enzyme Therapy
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Comparison of media conditions to
expand TIL-derived T-cell lines from patients with peritoneal
carcinomatosis including serum free, rIFN-g and
polyenzyme preparation
Kim Y.M, Zavadova E., Loercher A., Freedman R.S.
Dept of Obstet. & Gynecol, College of Medicine,
Univ. of Ulsan, Asan Medical Center, Seoul, Korea & Dept.
Of Gyn/Oncol. M.D. Anderson Cancer Center, Houston, TX
90th Annual Meeting, April 10 - 14, 1999 - Philadelphia,
published in American Association for Cancer Research
1999: Vol. 40, March 1999, Poster 1220.
491 KA (2-08-1) |
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The optimal culture conditions for the expansion
of TIL ex vivo have not been developed. In the interest
of patient safety, serum free conditions have been promoted
(J Immunol Meth 167:145-160, 1994). However, serum may contain
nutritional or other components that facilitate T cell line
growth. Objective: To develop improved culture conditions
for ex vivo expansion of TIL. Specimens & Methods:
We compared our serum free method of expansion of TIL from
19 patients under the following culture conditions: (a) +
5% FCS, (b) + 5% FCS + rIFN-g, (c) + rIFN-g, and (d) 5% FCS
+ rIFN-g + a polyenzyme preparation containing trypsin, chymotrypsin
and papain. Proteolytic enzymes are known to neutralize TGF-b
which inhibits lymphocyte proliferation. After 3 weeks in
culture TIL were examined for (i) fold of proliferation,
(ii) leukocyte cell surface antigens by FACS, (iii) cytokine
production by RT-PCR, and (iv) cytotoxicity against allogeneic
or autologous tumor cells. Results: (a) The fold of proliferation
of TIL grown in FCS was significantly better than cultures
without FCS. TIL with rIFN-g and polyenzyme preparation were
significantly better than with rIFN-g alone (p<0.001).
No differences were detected in expression of CD3+, TCRab+,
CD4+, or CD8+ T cells but a significant increase in expression
of CD56+ cells in the interferon group (p <0.001) and
the interferon group with enzymes (p <0.01) as compared
with medium alone. No differences were detected for in
vitro cytotoxicity to autologous or allogeneic cells.
TIL expanded in culture expressed mRNA for IL-2, IFN-g, GM-CSF,
and b-actin, but not IL-4 and IL-10. We conclude that culture
of TIL from patients with peritoneal carcinomatosis with
IFN-g and polyenzyme preparation significantly stimulate
the growth rate, but does not significantly change the phenotype
of lymphocytes. Supported in part by Betty Ann Asch Murray
fund for research in GYN/Med Oncol. |
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