Rosenberg concluded that adoptive immune therapy by means of 11-2 stimulated LAK cells could 
overcome defective cellular immunity to cancer (6, 18). In a variety of murine systems, he showed that 
the administration of 11-2 plus ex vivo amplified LAK cells led to marked regression of disseminated 
cancers (6,19,20) and cure of leukemia with (21) or without syngeneic marrow transplant (22). His 
studies also revealed the pleiotropic effects of 11-2, which include: endothelial cell antigen activation; 
stimulation of B cell proliferation; increased HLA class II (Dr) antigen expression on endothelial cells, 
tumor infiltrating lymphocytes (TIL), and melanoma cells; eosinophilia; and enhanced production and 
release of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) (9). Some of 
these may play important enhancing roles in the immune response to 11-2. 11-2 administration (with or 
without ex vivo generated LAK cells) in humans caused overall responses (including some durable 
complete remissions) in the range of 15-30% in patients with far-advanced, highly resistant neoplasms, 
such as melanoma and renal cancer (23-26) and relapsed acute leukemia (27). The high doses of 11-2 
used in the initial studies (even without reinfusion of cultured LAK cells) were associated with 
substantial toxicity, including capillary leak syndrome, severe hypotension, renal failure, liver injury 
and pulmonary insufficiency requiring intubation (26,28-31). This led to trials employing lower doses 
and/or continuous infusions (13,14,32-35) of 11-2. Toxicity, although reduced, still persisted, limiting 
the duration of therapy, and possibly the response rates as well. Attempts to achieve greater specificity 
and response by utilizing 11-2 plus TILs, harvested from the lymphocytes infiltrating some cancers, have 
thus far been technically difficult and associated with toxicity (2,9). The short half-life of 11-2, the 
toxicity of its systemic administration, and the rapid disappearance of effector cells after termination 
of 11-2 are major limiting features. 
The finding of tumor infiltrating lymphocytes in many tumors, or even in circulation, which 
however do not mount a effector response in vivo, could suggest that these lymphocytes may be 
specifically recognizing tumor cells but remain anergic or suppressed (36-38). According to current 
models, anergy in T-cells may be induced by the binding of ligands to the T-cell receptor variable 
region in the absence of second signals, which usually are provided by lymphokines (For review, 39). 
In support of such a hypothesis are the studies by Fearon et al. (40), Tepper et al. (41), Gansbacher 
et al. (42) demonstrating that nonimmunogenic tumors, upon transfection with IL2 or IL4, respectively, 
are rejected by syngeneic mice, whereas the normal (untransfected) tumor invariably kills the mice in 
a short period. These findings suggest that the production of lymphokines by the tumor can lead to its 
rejection. In other words a nonimmunogenic tumor seems to be rendered immunogenic upon lymphokine 
production by the tumor cells. Interestingly, tumor cell IL4 production caused tumor rejection even in 
nude mice suggesting the stimulation of non specific, inflammatory processes, involving mostly 
eosinophiles and monocytes. 
1.3 University of Miami Pre-CLinical Studies 
a. Effect of H2, 134, 116 and TNF transfection on the rejection of Lewis lung carcinoma 
We have completed a number of studies to determine whether the nonimmunogenic Lewis lung 
carcinoma (LLC) could be rendered immunogenic by transfection with the lymphokine cDNAs for 112, 
114, 116 and TNF and subsequent reinjection into murine syngeneic recipients (3,4 manuscript in 
appendix). LLC is a spontaneous lung tumor in C57B16 (43) which, when injected subcutaneously (10 6 
cells), will kill all recipients with a mean survival time of 34 days. No immune response is apparent 
and no infiltrating cells are detectable. 
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Recombinant DNA Research, Volume 18 
