Richard S. Schulof

Multivariate analysis of T-cell functional defects and circulating serum factors in Hodgkin's disease.

A comprehensive immunologic and serologic analysis was performed on 31 untreated patients with Hodgkins disease. Immune evaluations stressed T‐cell functional activity and included traditional parameters (PHA responsiveness and delayed hypersensitivity skin reactivity), as well as newer functional assays (T‐cell colony formation, chemotaxis, spontaneous and antibody‐dependent cytotoxicity, and concanavalin A‐induced suppressor cell activity (CISA)). Serum factors included ferritin, prostaglandins, zinc, copper, immune complexes, and thymic hormone activity. Every patient exhibited at least one T‐cell or serum abnormality. The greatest percentage of patients exhibited T‐cell defects in chemotaxis (85%), colony formation (81%), and PHA reactivity (64%). Immune defects were more common with advanced disease but were not related to absolute T‐cell or monocyte count, skin test anergy, or abnormalities of Tμ/Tγ cell proportions. Linear relationships were identified among abnormalities in the three assays employing mononuclear cells (PHA, colony formation, CISA) which may have reflected the inhibitory influence of monocytes present in the mononuclear cell preparations. Low serum zinc correlated with marked impairment of T‐cell chemotaxis. Elevated prostaglandins were associated with high PHA reactivity and with depressed colony formation. Our results indicate that many complex factors, including intrinsic T‐cell defects, contribute to the impaired immunity associated with Hodgkins disease.

In vitro effects of thymosin and lithium on lymphoproliferative responses of normal donors and HIV seropositive male homosexuals with AIDS-related complex

The in vitro effects of thymosin fraction 5 (TF5) and lithium chloride (LiCl) on the ability of peripheral blood mononuclear cells (PBMC) obtained from 37 normal male donors and 33 male patients with AIDS-related complex (ARC) to respond to alloantigenic stimulation (mixed leukocyte reaction, MLR) and to produce interleukin 2 (IL-2) in response to mitogens were studied. TF5 significantly increased MLR responses in normal donors (P less than 0.01) and in a group of 33 ARC patients with depressed cellular immunity (P less than 0.05). Similar effects were observed when LiCl was added to the MLR assays in both the normal and the ARC patient groups. Furthermore, TF5 and LiCl exhibited additive immunoenhancing properties. In 10 normal donors TF5 enhanced phytohemaggutinin (PHA)-induced IL-2 production as well as IL-2 production in response to pokeweed mitogen (PWM) (P less than 0.02). TF5-mediated enhancement of IL-2 production by PBMC obtained from ARC patients was observed in response to both mitogens, i.e., PHA and PWM. Additionally, LiCl increased PHA-induced IL-2 production in both normal subjects and ARC patients. LiCl and TF5 together had an additive effect in the enhancement of IL-2 production in both groups of subjects. Our data extend previous observations regarding the immunoregulatory activities of TF5 and LiCl and provide evidence that PBMC obtained from ARC patients have the potential to respond in vitro to these agents. The significance of these findings is discussed.

T‐cell abnormalities after mediastinal irradiation for lung cancer. The in vitro influence of synthetic thymosin alpha‐1

The effects of mediastinal irradiation (RT) on the numbers and functions of purified peripheral blood T‐lymphocytes from patients with locally advanced non‐small cell lung cancer were evaluated. The patients were candidates for a randomized trial to evaluate the immunorestorative properties of synthetic thymosin alpha‐1. Twenty‐one patients studied before RT did not exhibit any significant difference in T‐cell numbers or function compared to age‐matched healthy subjects. However, 41 patients studied within 1 week after completing RT exhibited significant depressions of E‐rosette‐forming cells at 4°C (E4°‐RFC)/mm3, E‐rosette‐forming cells at 29°C (E29°‐RFC)/mm3, OKT3/mm3, OKT4/mm3, and OKT8/mm3 (P = 0.0001); total T‐cell percentages (%OKT3, P = 0.01); and T‐cell proliferative responses in mixed lymphocyte cultures (MLR) (P = 0.01) and to the mitogen phytohe‐magglutinin under suboptimal conditions (P ≤ 0.03). Nine patients studied before and after RT showed a significant increase in OKT4/OKT8 (P = 0.01) following RT. A short‐term in vitro incubation with thymosin alpha‐1 could enhance MLR of T‐cells in 12 of 27 patients with post‐RT abnormalities. In 13 patients who were treated with placebo, the RT‐induced depression of T‐cell numbers and function persisted for at least 3 to 4 months. In addition, in 12 patients progressive decreases developed in %E4°‐RFC, %OKT3, %OKT4, and OKT4/OKT8. which always preceded clinical relapse. This study indicates that mediastinal RT results in prolonged depletion of circulating T‐cells, alterations of T‐cell subset proportions, and intrinsic T‐cell functional deficiencies. This patient population provides a uniformly immunosuppressed group of subjects with which to evaluate the immunorestorative effects of thymosin alpha‐1 or other biologic response modifiers.

Thymosin in the early diagnosis and treatment of high risk homosexuals and hemophiliacs with AIDS-like immune dysfunction.

The adnormal levels of thymosin alpha 1 in acquired immunodeficiency syndrome (AIDS) patients, depressed T-cell function, thymus pathology, and the restoration of T-cell function by thymosin fraction 5 (TF5) lend support to the hypothesis that the thymus plays a central role in AIDS. The thymosin alpha 1 assay may provide a means of identifying symptomatic carriers of AIDS. This paper summarizes the current status of diagnostic studies with thymosin alpha 1 in AIDS and reports the 1st clinical trial with thymosin in subjects with AIDS-like immune dysfunction. Serum samples from intravenous drug abusers, homosexuals, and Haitians with AIDS have revealed thymosin alpha 1 levels at least 2 standard deviations from the mean of controls without AIDS. Preliminary data from a pilot study in homosexuals and hemophiliacs at high risk for AIDS suggest that the administration of TF5 may be effective in reconstituting some T-cell mediated specific immune functions, including cell-medicated lympholysis (CML) and the mixed lymphocyte response (MLR), and enhancing the lectin-induced production of T-cell growth factor. On the other hand, TF5 has failed to have any effects on the T4/T8 ratio, absolute lymphocyte counts, or natural killer cell activity.

Phase II trial of subcutaneous interleukin-2, subcutaneous interferon-α, 5-fluorouracil and cis-retinoic acid in the treatment of renal cell carcinoma: Final results of cancer biotherapy research group 94-10

BACKGROUND The treatment of metastatic renal cell cancer remains unsatisfactory despite encouraging results with biotherapy. Pilot studies from other investigators have suggested that combining cis-retinoic acid and 5-fluorouracil (5FU) with interleukin-2 (IL-2) and interferon-alpha (IFN) may improve outcomes for such patients. METHODS Eligible patients had metastatic renal cell cancer, were in good medical condition, and had not been treated previously with more than two of the study agents. A 56-day treatment cycle consisted of: interferon-alpha 2a 3.0 mu/m2 s.c. Monday, Wednesday, and Friday weeks 1-8, interleukin-2 11 mu/m2 s.c. Tuesday, Thursday and Saturday of weeks 1-4, cis-retinoic acid 1 mg/kg p.o. daily weeks 1-8, and 5-FU 750 mg/m2 i.v. weekly during weeks 5-8. Patients were evaluated for tumor response every 8 weeks, and in the absence of tumor progression, patients could receive treatment for up to one year. Survival was determined from the first date of treatment. RESULTS The 58 renal cell carcinoma patients included 41 men and 17 women, with a median age of 57 years with a range of 28-85 who were enrolled between October 1994 and July 1997. Thirty-seven percent were asymptomatic when treatment was initiated. Sites of disease at study entry included 62% lung, 34% bone, 31% lymph node, 22% kidney, 16% liver and 10% adrenal. There were only three patients with significant tumor responses (one complete, two partial) for a response rate of 5% (0-11% 95% CI) based on intent-to-treat analysis, and 6% (0-12%, 95% CI) for the 53 patients who were evaluable for response. The response rate among evaluable nephrectomized patients who had received no prior radiation or systemic treatment was 3/25 (12%). The median failure-free survival was 2.8 months; median overall survival was 10.9 months. The 1-year survival rate was 50% and 2-year survival rate was 33%. The most frequent toxicities were fatigue-81% (26% grade 3 or 4), nausea/vomiting-59%, and leukopenia/neutropenia 57% (16% grade 3 or 4). CONCLUSION Despite a disappointing objective response rate, survival in these patients who were treated entirely as outpatients was similar to that seen in our earlier trials of inpatient, intermediate dose continuous infusion IL-2-based therapy.

Inverse correlation between age related abnormalities of T-cell immunity and circulating thymosin α1 levels in haemophilia A

T‐cell immunity and serum levels of thymosin α1, β2‐microglobulin, circulating immune complexes, serum immunoglobulin levels, antibodies to hepatitis surface or core antigen, and to cytomegalovirus, and Epstein‐Barr virus were investigated in 51 patients with haemophilia A ranging in age from 2 to 52 years. All patients had received commercial U.S. lyophilized concentrates of antihaemophilic factor (AHF). The mean helper/cytotoxic‐suppressor (0KT4/0KT8) ratio of 11 pre‐adolescents (1‐6 ±0‐4 SE) was not significantly different from that of age matched normal controls. In contrast, the mean 0KT4/0KT8 ratios of 13 adolescent (1‐2 ±0‐2 SE) and 23 adult (0‐8 ±0‐1 SE) haemophiliacs were significantly reduced. Abnormalities of lymphocyte mitogenic responses were found only in adult haemophiliacs. Nine individuals treated with commercial U.S. prothrombin complex concentrates for antibodies directed against AHF or for haemophilia B had normal mean OKT4/OKT8 values. The mean serum thymosin α1 levels for each age category was similar to that of age matched controls; however, regression analysis revealed a significant relationship between elevated thymosin α1 levels and decreased OKT4/OKT8 ratios in adult haemophiliacs (P= 0‐012). Although the mean serum level of β2‐microglobulin was significantly increased in the adult haemophiliac group, there was no correlation between OKT4/OKT8 ratios and any of the other serologic parameters studied.

Clinical virologic and immunologic effects of combination therapy with ribavirin and isoprinosine in hiv infected homosexual men

We evaluated the clinical, immunologic, and virologic effects of oral treatment with ribavirin and isoprinosine for up to 3 months in asymptomatic, HIV-culture-positive homosexual men. Fifteen consecutive men received isoprinosine 4 g/day (1 g q.i.d.), and 800 (9 men) or 1,200 mg/day (6 men) of ribavirin. Five men in each ribavirin dosage group completed at least 2 months of treatment. No unexpected toxicities were observed. Eight minor HIV-related events occurred in six men while on study. All men remained HIV-positive, and time to positive culture decreased by at least 4 days in three men from each treatment group. Serum p24 levels did not change in two men who were p24 antigenemic and received 800 mg/day of ribavirin. Treatment was associated with a generalized lymphopenia affecting all lymphocyte subsets including CD4, which was partially reversible 1 month after stopping treatment. Most of the men remained anergic on DTHS skin testing. No improvements were noted in in vitro lymphoproliferative responses to antigens or in NK cell activity (which decreased significantly in the 1,200 mg/day ribavirin group). Although well tolerated at the doses employed, the combination of ribavirin and isoprinosine produced an unexpected generalized lymphopenia and did not exhibit HIV-suppressive or immunorestorative effects.

Thymosin in the Staging and Treatment of HLTV-III Positive Homosexuals and Hemophiliacs with AIDS-Related Immune Dysfunction

The paralysis and destruction of the thymic dependent immune system has become the hallmark of the acquired immune deficiency syndrome (AIDS) (1). Clinical symptoms of the disease usually include a progressive decline in essentially all thymus mediated immune functions, an increase in susceptibility to pneumocystis carinii pneumonia and other opportunistic infections, and an increase in the incidence of Kaposi’s sarcoma and a number of lymphoid tumors (2–4). The current prognosis for patients with AIDS is dismal. Over 7,000 cases of AIDS have now been reported to the Center for Disease Control and over 70% of the patients have died. Immunological abnormalities which accompany the disease include a decline in helper T-cell number and function, a decline in lymphocyte reactivity to mitogens and antigens, decreased production of lymphokines (such as T-cell growth factor, IL-2, and interferon), and an increase in thymosin α1 like cross-reacting material in serum. The presence of antibodies to a number of viruses has been documented and a recently described retrovirus designated as HTLV-III has been implicated as the cause of AIDS (1,5,6). To date AIDS has been confined primarily to individuals who fall into a relatively small number of risk groups. They include homosexuals (78 %), IV drug addicts (14 %), Haitians (3 %), and hemophiliacs (1 %) (1). The remaining small number of individuals (4 %) are generally found to be associated in some way with the defined risk groups.

Continuous-infusion floxuridine and alpha interferon in metastatic renal cancer: a national biotherapy study group phase II study.

Eighteen patients with advanced renal cancer were treated with 0.15 mg/kg/day floxuridine by continuous intravenous infusion for 14 days with 3 million IU/m2/day alpha interferon subcutaneously three times weekly. Treatment cycles were repeated every 28 days. Floxuridine dosages were escalated to a maximum of 0.2 mg/kg/day and alpha interferon dosages were escalated to a maximum of 6 million IU/m2/day depending on patient tolerability. A total of 49 treatment courses were administered with a median of 2.7 courses per patient. Of 14 assessable patients, there were no complete or partial responses. Eight patients (57%) had stabilization of disease. The median survival for patients with stable disease was 20.9 months and for all 18 patients was 7.2 months. Grades 3 and 4 toxicities included diarrhea (44%), nausea/vomiting (28%), mucositis (11%), fever (22%), and fatigue (50%). Dose-limiting toxicities were primarily gastrointestinal symptoms. There were no treatment-related deaths. This combination in the dose schedule used did not result in any significant objective tumor response but was associated with considerable toxicity.

An Evaluation of Two Different Schedules of Synthetic Thymosin α1 Administration in Patients with Lung Cancer

Thymosin α1 (molecular weight 3108) is one of the many active polypeptides isolated from thymosin fraction 5 (TF5) (Low and Goldstein, 1979; Goldstein et al., 1982). α1 was initially purified from extracts of calf thymus glands. Biologically active α1 has now been successfully synthesized by classical solution (Birr and Stollenwerk, 1979; Wang et al., 1978), solid-phase (Wang et al., 1980; Folkers, et al., 1980) and recombinant DNA procedures (Wetzel et al., 1980).