GB2194146A - Treatment of retroviral infections - Google Patents

Abstract

Retroviral infections e.g. Aids or Aids related complex, are prevented or treated by administration of tumor necrosis factor, optionally with an inteferon; and preferably by continuous infusion of tumor necrosis factor and interferon.

Description

SPECIFICATION Compositions for the prophylaxis and treatment of retroviral infections This invention relates to the prevention and treatment of infections by retroviruses. In particular, it is concerned with the therapy of retroviral infections leading to immunological defects, in particular those infections thought to be responsible for acquired immune deficiency syndrome (AIDS).

AIDS is a transmissible deficiency of cellular immunity characterized by opportunistic infections and certain malignancies, notably Pneumocystis carinii pneumonia and Kaposi's sarcoma, in patients without another recognized cause for contracting these rare diseases (1-3). AIDS is manifsted by a profound lymphopenia, a generalized cutaneous anergy and a markedly reduced proliferative response to mitogens, antigens and aliogeneic cells, seeming to result from depletion of the OKT4+ T-lymphocyte subset (4). While humoral immunity is relatively unafected, there is increasing evidence for a hyperactive B-cell proliferative response which may be related causally to the high incidence of B-lymphoma in AIDS patients (5,6).In addition to the fully developed syndrome, an epidemic of a related disease, AIDS-related complex (ARC), has appeared, characterized by generalized chronic lymphadenopathy. This syndrome shares many of the epidemiological features and immune abnormalities and often precedes the clinical manifestations of AIDS.

Recent evidence has implicated strongly a lymphocytotropic retrovirus as the primary aetiological agent of AIDS and the AIDS-related complex. Lymphadenopathy-associated virus (LAV) was isolated initially from cultured lymph-node T cells of patients with lymphadenopathy and AIDS as well as an AIDS patient and an asymptomatic sibling, both with haemophilia B (7-9). A similar virus, designated human T-lymphotrophic virus type Ill (HTLV-III), has been isolated from a large number of AIDS and ARC patient blood samples by co-cultivation with the permissive T-cell line H9 (10, 11). LAV and HTLV-III, as well as related retroviruses isolated recently from AIDS patients (12, 13), share several important characteristics.Viral replication occurs in the OKT4+ T-lymphocyte population in vivo and in vitro and is associated with impaired cell proliferation and the appearance of cytopathic effects (8,10,14). The virus has a Mg21-dependent reverse transcriptase, exhibits a dense cylindrical core morphology similar to type D retroviruses (8,13,15) and is recognized by antibodies found in the sera of virtually all AIDS and ARC patients (8,13,16-21). HTLV-III and LAV now are believed to be strains of the same virus, which has been given the name human immunodeficiency virus (HIV).

Immune function in AIDS or ARC afflicted patients is severely compromised. However, the precise nature of the abnormalities induced by HIV continue to be under study. Of particular interest was the effect of HIV infection on the production of'lymphokines by the immune cells of such patients well as their responses to oxogenously administered lymphokines.

Of 16 AIDS patients tested for T lymphocyte capacity to secrete macrophage-activating products, including gamma interferon, 14 failed to generate active lymphokines and 13-14 completely failed to secrete gamma interferon. Furthermore, macrophages from patients with AIDS showed enhanced antimicrobial activity when incubated in vitro with gamma interferon, thus raising the possibility that such cells may respond in vivo to exogenously administered gamma interferon (22, 24).

Lymphocytes from AIDS patients also have been reported to be deficient in their ability to produce interleukin-2, a lymphokine having a role in the proliferation and differentiation of T lymphocytes and which stimuiates gamma interferon production (23).

Finally, gamma interferon was known to exert a diect antiviral effect on vesicular stomatitis virus (VSV) infected cells and to potentiate the effect of a cytokine on VSV infected cells (25).

This cytokine was tenatively identified by these authors as the polypeptide referred to herein as tumor necrosis factor-a. See also Eifel et a/., "Cell. Immun." 47: 197-203 (1979). Immunosupplementation therapy for AIDS therefore appeared to be a promising opportunity, and clinical studies with patients were instituted.

Despite the expectations for lymphokine therapy of AIDS patients, in vivo results have been disappointing. Contrary to the results encountered in in vitro experiments, a prolonged course of intravenous recombinant gamma interferon treatment appeared to inhibit rather than enhance monocyte respiratory burst function (26), and intravenous treatment of AIDS patients with varying doses of interleukin-2 or gamma interferon led to the conclusion that in vivo lymphokine therapy was not of significant value in the treatment of patients with AIDS (27).In fact, by the time of the invention herein, a school of thought had developed that the immune stimulation of reconstruction approach to AIDS therapy was actuaily ''dangerous" because cell activation by lymphokines was thought to contribute to virus spreading and T4 cell depletion, and thus would accelerate progression of the disease (28). In June 1986, Yamamoto et al. (29) disclosed that human alpha and beta (but not gamma) interferons suppressed the in vitro replication of HIV strains, but when exposure to the interferons ceased the viral production of infected cells was enhanced. This further suggested that immune therapy of AIDS would be counterproductive.

Finally, it has been suggested that lyphotoxin is produced by HIV-infected T cells by virallyinduced transactivation of the T cells' endogenous lymphotoxin gene, thus leading to secretion of autotoxic quantities of lymphotoxin (35).

Tumor necrosis factors are polypeptides produced by mitogen-stimulated macrophages (TNF-a) or lymphocytes (TNF-ss) which are cytotoxic for certain malignantly transformed cells. TNF-a has been suggested to be responsible for wasting and cachexia in patients with cancer of severe infections (34), and passive immunization against TNF-a was reported to protect mice against the lethal effect of endotoxin (30). The anti-tumor effect of TNF-a is known to be synergistically potentiated by interferons, and the anti-tumor effect of TNF-ss is similarly potentiated by interferon gamma. The anti-tumor effect of TNF-a and TNF-ss in admixture, however, are only additive, as are the antiviral effects of interferons alpha and beta.

It is desirable to employ immunotherapy in the successful treatment of persons infected with retroviruses, in particular lymphocytotropic viruses such as HIV, and also to treat those at risk for active retroviral infection, e.g. persons harboring latent retrovirus and provide immunotherapeutic prophylaxis against retroviral infections.

It has been discovered that administration of a therapeutically effective amount of a tumor necrosis factor alone or, preferably, in combination with an interferon, can confer protection on persons at risk to retroviral infection and kills retrovirally infected cells. While gamma interferon alone exerts little or no protective activity against retroviral infection nor is gamma interferon significantly cytotoxic to retrovirally infected cells, and TNF alone is only modestly active at high concentrations, the combination of these two agents is dramatically potent. This phenomenon is observed in vivo, despite the deranged condition of the immune systems of retroviraily-infected patients. This result was particularly surprising because TNF was not known to be deficient in such patients.

Brief Description of the Figures Figure 1 is a Northern gel showing the dramatic reduction in HIV mRNA in HIV-infected HuT78 cells after pretreatment with, TNF-a and IFN-y as compared to control cells that were not pretreated.

Figure 2 demonstrates the antiviral protective effect of catalase in combination with TNF-a and/or IFN-r.

Detailed Description Retroviruses are defined as viruses containing single or double stranded RNA. These viruses replicate by harnessing the cellular metabolism of permissive hosts to reverse transcribe the RNA genetic material of the virus. The large amounts of DNA so produced are translated into HIV protein and RNA for the assembly of progeny virions. Examples of such viruses include the socalled "slow" or lentiviruses and the T-cell leukemia viruses such as HTLV-I and HTLV-II, but most preferably are the HIV strains associated with AIDS.

The tumor necrosis factors useful herein include TNF-a and TNF-fl. The former is described in copending U.S.S.N. 881,311, filed July 2, 1986, together with methods for its synthesis in recombinant cell culture. Similarly, the latter (previously called lymphotoxin) and suitable recombinant synthesis methods are described in copending U.S.S.N. 616,502, filed May 31, 1984 and U.S.S.N. 732,312, filed May 9, 1985. The TNF-a and TNF-ss described in these applications include cytotoxic amino acid sequence and glycosylation variants which also are used herein. Of course, TNF-a or TNF-ss from non-recombinant sources are also useful in the method of this invention.

TNF-a or TNF-ss are used alone or in admixture with one another in proportions empirically determined to exert the most effective clinical response. TNF is not species specific, so TNFs from other animal species, e.g. porcine or bovine, are useful herein. The preferred TNF is mature human TNF-a from recombinant microbial cell culture. The TNF ordinariiy will have a cytolytic activity on susceptible L-929 murine cells of greater than about 1 x 106 units/mg, wherein a unit is defined as set forth in the above-described patent applications, the disclosures- of which are incorporated by reference.

Interferons are well known. In nature they comprise the ss, y and about 20 different interferona subtypes. Their most relevant characteristic for the purposes herein is that they be capable of protecting cells in vitro and in vivo from viral infection. The interferons used in the process or composition of this invention typically are interferons-a, ss and/or y. Interferons produced in recombinant cell culture, from natural isolates or by stable untransformed cell lines are satisfactory for use herein, as are interferon amino acid sequence or glycosylation variants (including unglycosylated forms) so long as they exhibit anti-viral activity. Interferon-y should be of the same animal species for which therapy is intended because interfernn-y is known to be species specific. The interferons desirably are substantially homogeneous and will have a specific activity in excess of about 1 x 106 International Units/mg.

The compositions herein include a pharmaceutically acceptable vehicle such as those heretofore used in the therapeutic administration of interferons, TNF or LT, e.g. physiological saline or 5% dextrose, together with conventional stabilizers and/or excipients such as human serum albumin or mannitol. The compositions are provided lyophiiized or in the form of sterile aqueous solutions.

Several variables will be taken into account by the ordinary artisan in determining the proportions of interferons a, ss or y and of TNF-a or TNF-ss, the net proportion of interferon to TNF, the concentration of interferon and TNF in the therapeutic compositions and the dosages to be administered on a Kg basis. Therapeutic variables also include the animal species to be treated, the administration route, and the clinical condition of the patient (including the stage and degree of retroviral and/or adventitious organism infection, if any, present at the commencement of treatment). Doses of interferon ranging about from 1 to 50 ,ug/m2 are suitable initial dosing levels. The tolerated dose may not exceed about 25 Ag/m2.

The TNF and interferon doses are administered together or separately. If the latter, the interferon should be administered first and the TNF thereafter within 24 hours. It is within the scope of this invention to administer the TNF and interferon in multiple cycles, depending upon the clinical response of the patient. This approach to the therapy will be effective in attacking latently-infected cells entering the active phase of infection, whether due to exogenously administered T cell mitogens or adventitious infections that lead to T cell activation.

Since the treatment with TNF and interferon will lyse virally-infected cells and may result in the relese of infective virus, it is advantageous in the course of therapy to administer substances capable of neutralizing further viral infectivity. This can be accomplished by several methods. One can administer antibodies such as monoclonal or polyclonal anti-retroviral antibodies during the course of therapy, preferably at the same time as TNF is administered. Alternatively, immunologically competent patients can be vaccinated against the retrovirus in order to actively induce neutralizing antibody. A suitable vaccine for this purpose contains the HIV gp120 env and is described in U.S.S.N. 861,016, filed May 8, 1986. This vaccine is disciosed to induce HIV neutralizing antibodies, which in turn can be used in the passive immunization strategy described above.Other agents that interdict the potential infectivity of released viruses also can be administered together with the TNF, for example gp120 env or fragments thereof which bind to the cell surface receptor ordinarily recognized by the retrovirus in question (in the case of HIV, the OKT4' cell surface marker present on helper T cells) and thus competitively inhibit viral adhesion to target cell surfaces.

The synergistic antiviral and anti-tumor activity of TNF and/or an interferon is further potentiated by including in the treatment regimen and/or compositions a therapeutically effective amount of an oxygen free-radical scavenger (including oxygen protective enzymes and peroxidatively active substance). Such substances, including catalase, superoxide demutase, peroxidase or chloroperoxidase, greatly enhance the activity of TNF and interferon. Such enzymes are respectively known to catalyze the breakdown of H202 to water and oxygen or H202+HO-R OH2H2O+O-R-O, although at present the mechanism(s) by which such agents potentiate TNF and interferon are unknown. Catalase is widely available commercially and can be obtained- from human tissues such as red blood cells. Peroxidase is commonly available from horseradish.The proportion of peroxidatively active substance to TNF and interferon optionally will be about from 1000:1:0.1 to 100:50:25, but the dosages and proportions necessarily will be adjusted to reflect the clinical condition of the patient and administration route, among other variables known to those skilled in the art.

The TNF and interferons are administered by the same or separate routes, for example by intravenous, intranasal or intramuscular administration. Either or both components can be administered from sustained reiease compositions, for example as polylactide or polyhydroxylbutyrate implants or liposomes such as are described in EP 17,2007A, or by continuous infusion. At the present time it is preferred to infuse the TNF and interferon intravenously in the dosages described above.

In order to avoid side effects from the combined therapy when high doses are used, the TNF and interferon can be employed in an extracorporeal treatment regimen heretofore referred to as adoptive immunotherapy (U.S.S.N.s 763,657 and 743,570, filed August 8, 1985 and June 11, 1985, respectively). In these methods the peripheral blood mononuclear or lymphocytic cells of a patient are separated from the blood in an extracorporeal plasmapheresis cycle, treated with interleukin-2 or interferon, and then reinfused into the patient. The patient's immune responses against malignancies were greatly stimulated by this procedure. For the treatment of retroviral infections, the same general procedure is employed except that the peripheral monocytes and/or lymphocytes are incubated in the presence of TNF or TNF and interferon. Virally-infected cells are killed by TNF or the combination of agents.The extracorporeal cells also can be incubated with a killer cell activating agent such as a lymph cell mitogen (e.g., phytohemagglutinin) and/or interleukin-2. The cells then are washed and resuspended in an infusion medium for reinfusion into the same patient from whom the cells were first obtained. The reinfusion can be followed up by administration of TNF and interferon as is otherwise provided herein. The optimal amounts of TNF or TNF and interferon, and the optimum conditions for extracorporeal treatment of the patients' lymph cells are routinely determined by assaying the patients' viral titer and assessing improvements in the patients' clinical condition.

Persons that are candidates for treatment in accordance with this invention are those at risk for exposure to retroviral infection or who exhibit signs of actual exposure to such an infection.

Those at risk include members of high risk groups such as homosexuals, intravenous drug users and those who have received transfusions or other products made from blood not screened for HIV antibodies. Evidence of exposure to retrovirus includes sero-conversion to antibodies against HIV, positive serum assays for HIV, symptoms associated with AIDS-related complex (ARC) or frank AIDS. AIDS patients may or may not be diagnosed for Kaposi's sarcoma or other malignancies, adventitious microbial infections such as Pneumocystis carinii or thrush, neural impairment of cachexia (wasting).

The invention will be more fully understood in the light of the following examples. The interferons were the products of recombinant bacterial cell culture and were purified to a specific activity of about 108 units/mg. TNF-a and TNF-ss also were recombinant, and had a specific activity of about 5x107 units/mg. Ali literature citations are hereby incorporated by reference.

Example 1 Prophylaxis of HIV Infection The OKT4±positive human T-cell leukemia cell lines H9, HuT78 and U937 were cultured in suspension with RPMI-1640 medium at 37"C. Cultures were centrifuged to separate cells from the medium and the cells then resuspended in fresh RPMl-1640 at a density of 1 x 106/ml.

Sufficient TNF-a and gamma interferon were added to the resuspended culture to produce 0.1 ,ag/ml TNF-a and 0.1 Xlg/ml gamma interferon. The combination of TNF-a and gamma interferon was not toxic to uninfected HuT78 and H9 cells. The culture then was incubated at 37"C for 24 hours, after which 1 x 106 cpm units of HlV/ml was added. The cpm units were determined by reverse transcriptase activity (31); each unit is calibrated to represent the reverse transcriptase activity of one virion. After 3 days of further incubation at 37"C the cells were screened for viral antigens by indirect immunofluorescence using a murine monoclonal antibody against the p24 (core) protein of HIV and labelled goat anti-mouse immunoglobulin (32).The results of replicate experiments are shown below as the percentage of cells which contain HIV core protein.

Table 1 Agent % of Cells Infected HuT78 H9 U937 1 2 1 2 1 2 Control 54 68 48 36 11 14 TNF-a 27 34 32 18 7 8 gamma interferon 63 51 42 41 12 11 TNF-a and gamma interferon 1 8 4 3 2 1 These results demonstrate that combination therapy with TNF and interferon is highly effective in preventing HIV infection of otherwise susceptible T cells.

Example 2 Treatment of HlV-lnfected Cells 1 x 105 H9 and RPMI-1788 lymphoblastoid cells/ml of RPMI-1640 medium were exoposed to 1x103 cpm of HlV/ml in the presence of 1 ,ug/ml polybrene in order to enhance the percentage of cells infected by HIV. These cells were cultured for about 30 days, during which the culture medium was replaced every 3-5 days and cells subcultured approximately once a week. Sufficient TNF-a and gamma interferon were substantially simultaneously added to cultures containing 1 x 106 cells/ml in order to establish concentrations as shown in Table 2 below. Combinations of TNF-a and gamma interferon at these concentrations were not cycotoxic towards uninfected H9 and RPMI-1788 cells. The cultures were incubated at 37"C for 3 days and then assayed for the percentage of viable cells by trypan blue staining. The results of replicate experiments are shown in Table 2 (ND=not done).

Table 2 Agent Concentration % Viable Cells H9 1788 1 2 1 2 Control - 89 76 95 86 TNF-a 1.0 ng/ml ND ND 90 81 0.1 ,ug/ml 85 74 81 76 Gamma 1.0 ng/ml ND ND 83 84 interferon 0.1 ,ug/ml 85 74 80 82 TNF-a and Gamma 1.0 ng/ml ND ND 68 72 interferon 0.1 g/mi 50 42 34 53 These results demonstrate concentration-dependent synergistic killing of virally-infected cells by combinations of TNF and interferon and, to a lesser degree, TNF-a alone. This, combined with the protective effect of TNF or the combinations for uninfected cells, demonstrates the value of TNF or combination therapy in the treatment of retroviral infections.

Example 3 Reduction in HIV Replication by TNF and Combination Therapy HuT78 cells were treated with 0.1 ,ag/ml each of TNF-a and gamma interferon and then infected with HIV as described in Example 1. After 2 days of incubation at 37"C total RNA was extracted from the HlV-infected cells as foliows. The cells were washed with PBS and resuspended in 0.35 ml TSM buffer (10 mM Tris pH 7.5, 0.15 M NaCI, 2mM MgCl2) containing 0.5% NP-40 and 17 Cil VRC (BRL) at about OCC. After 3-5 min. nuclei were centrifuged free of the lysed cells.The supernatant containing mRNA was added to 0.35 ml of TSE buffer (10 mM Tris pH 7.5, 0.15 M NaCI, 5 mM EDTA) containing 1% SDS and extracted 3x with 0.7 ml of phenol:chloroform:iso-amylalcohol (24:24:1). The phenol was equilibrated with water and 0.1% 8-OH-quinoline. RNA was precipitated from the aqueous layer with ETOH and sodium acetate.

Poly(A) RNA was prepared by oligo(dT) cellulose electrophoresis (36). Northern hybridization using 1 iLg RNA/lane was performed as described (37). The results, shown in Fig. 1, demonstrate that TNF-a pretreatment was able to suppress the appearance of HIV RNA in host cells, but that far less RNA appeared .when TNF and interferon were used together. These results are consistent with the infectivity data shown in Table 1. No change in actin mRNA (a structural protein) was observed with or without TNF and interferon. treatment, demonstrating that the effect of TNF-a and interferon was targeted to the virus rather than cell growth in general.

Example 4 Protocol for AIDS or ARC Patient Therapy Males seropositive for HIV antibody and having blood which is in vitro cell culture positive for demonstrable HIV titer are enrolled. These patients generally present with symptpmology consistent with AIDS or ARC, and were seroconverted for HIV antibody. The treatment group is divided into cohorts based on the following treatment variables: 1. Simultaneous or sequential TNF and interferon treatment.

2. Intravenous or intramuscular injection and, in the case of intravenous injection, by bolus or continuously by infusion (1, 2, 5 and 10 days).

3. Dosage regimen over hours, starting at 1 ,eig/m2 of TNF, and 1 Ag/m2 of interferon, and increasing to 5, 10, 25, 50... g/m2 of each agent as tolerated.

4. Ratios of TNF to interferon: 1:100 to 100:1 5. Repeat cycles of treatments: 1 to 5, with intermediate treatment suspensions of 1, 2, 5 or 10 days.

6. Selection and proportions of interferon types: a and y. 1:10-10:1; ss and 7, 1:10-10:1; 7, a or fi alone, 1:1:1 to 10:1:1 to 1:5:5.

A suitable starting regimen will comprise treating ARC patients with a combination of TNF-a, gamma interferon and alpha interferon. The combination is administered by continuous infusion using an intravenous infusion pump calibrated to deliver a dosage of 1-10 Ag/m2/24 hours TNFa, 1-10 ,ag/m2/24 hours for gamma interferon and 1-10 ,ug/m2/24 hours for alpha interferon.

Patients may be escalated to higher doses as tolerated. This infusion is continued for one week.

Patients are rested for another week and the infusion repeated. Side effects such as fever, chills and the like are treated by conventional means or by reducing the dosage. When anti-HIV antibody or gp120 env are employed as antiviral agents in combination with the above, the antibody or env polypeptide are present in a dosage calculated to be capable of sequestering virions released by the combined therapy. This will depend on the affinity of antibody for virion and its neutralizing titer as well as the viral titer of the patient. Determination of suitable dosages will be within the skill of the routineer.

The clinical condition and viral infectivity titers of the patients are monitored during and after the treatment protocol. Consistent with the in vitro studies in Examples 1-3, the immune competence and viral titer of a statistically significant proportion of the patients improved as a result of the treatment.

Example 5 Catalase Potentiates the Antiviral Activity of Interferon-y and/or TNF-a A549 cells were seeded at 2x 104/welI in 96-well flat-bottom trays (Falcon Plastics) for 24 hr prior to incubation with test samples. The test samples were as shown in Fig. 2 ("CAT" is catalase). The samples otherwise contained Dulbecco modified Eagle's (DME) medium supplemented with 5 percent heat-inactivated fetal calf serum (FCS), glutamine (2 mM), penicillin (100 U/ml), and streptomycin (100 ,ug/ml). After 18 hr of incubation at 370C, culture supernatants were replaced with fresh DME medium free of interferon, TNF or catalse but containing 2 percent foetal calf serum and EMC virus at a multiplicity of infection ("MOI", the ratio of infectious virus/cell) of 1.The cytopathic effect (CPE) was determined by staining the viable cells with crystal violet and the titer was quantitatively monitored using a microelisa autoreader (MR580, Dynatech) and further confirmed visually. The CPE antiviral titer is expressed as the reciprocal of the dilution found to inhibit 50 percent of the cell cytopathy and was standardized against the international reference sample of human interferon-y (No. Gg 23-go 1-530).

As shown in Fig. 2, catalase dramatically and synergistically enhanced the anti-viral effect of interferon-y and TNF-a. This effect is not unique to EMC virus. It has been observed with a variety of permissive cells and viruses.

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Claims (30)

1. A combination, for simultaneous or separate administration, of a tumor necrosis factor and an interferon, for use in the treatment of prophylaxis of retroviral infections.

2. The use of a tumor necrosis factor, or a tumor necrosis factor and an interferon, in the manufacture of a medicament for the treatment or prophylaxis of retroviral infections.

3. The composition or use of claim 1 or claim 2 wherein the tumor necrosis factor is tumor necrosis factor or tumor necrosis factor.

4. The composition or use of claim 3 wherein the tumor necrosis factor is tumor necrosis factor.

5. The composition or use of claim 1 or claim 2 wherein the interferon is interferon a, ss or 7.

6. The composition or use of claim 1 or claim 2 wherein the interferon is interferon-y.

7. The composition or use of claim 1 or claim 2 wherein the patient is infected with HIV.

8. The composition or use of claim 7 for patients afflicted with AiDS-Reiated Complex.

9. The composition or use of claim 1 or claim 2 for patients free of any detectable malignancy.

10. The-composition or use of claim 1 or claim 2 wherein the tumor necrosis factor or tumor necrosis factor and interferon are for administration ex vivo in the course of adoptive immunotherapy.

11. The composition or use of claim 10 wherein the tumor necrosis factor or tumor necrosis factor and interferon are for administration in addition intravenously or intramuscularly after the course of ex vivo adoptive immunotherapy.

12. The composition or use of claim 1 or claim 2 wherein interferon is adminstrable prior to tumor necrosis factor.

13. The composition of use of claim 11 wherein the interferon is a mixture of gamma interferon plus alpha or beta interferon.

14. The composition or use of claim 12 wherein the interferon is gamma interferon.

15. The composition or use of claim 11 wherein the dosages of tumor necrosis factor and interferon are each about from 1 to 25 zg/m2/24 hours by continuous intravenous infusion.

16. The composition or use of claim 11 further comprising for administration antibody capable of neutralizing viral infectivity.

17. The composition or use of claim 11 wherein the administration of tumor necrosis factor and interferon is repeatable through about from 1 to 5 cycles.

18. The composition or use of claim 6 further comprising means for immunizing the patient against HIV prior to administration of tumor necrosis factor or tumor necrosis factor and an interferon.

19. The composition or use of claim 11 wherein the tumor necrosis factor and interferon are administrable by continuous intravenous infusion for about from 1 to 5 days.

20. The composition or use of claim 1 further comprising for administration an anti-viral therapeutic dose of a physiologically acceptable oxygen free-radical scavenger substance.

21. The composition or use of claim 20 wherein the scavenger substance is catalase.

22. The composition or use of claim 21 wherein the scavenger substance is human erythrocyte catalase.

23. A composition comprising a tumor necrosis factor, an interferon and an oxygen freeradical scavenger substance.

24. A composition of claim 23 wherein the substance is a peroxidatively active enzyme.

25. A composition of claim 23 wherein the enzyme is human erythrocyte catalase.

26. A composition of claim 23 further comprising a substance capable of preventing (a) replication of a retrovirus or (b) retroviral binding to a cell surface receptor.

27. A composition comprising a tumor necrosis factor, an interferon and a substance capable of preventing the (a) replication of a retrovirus or (b) retroviral binding to a cell surface receptor.

28. The composition of claim 20 wherein the substance capable of preventing retroviral binding to a cell surface receptor is an antibody directed against an envelope polypeptide of the retrovirus.

29. A composition for use in the treatment or prophylaxis of retroviral infections, substantially as described herein.

30. The use of a tumor necrosis factor, optionally in combination with an interferon, in the manufacture of a medicament for the treatment or prophylaxis of retroviral infections, substantially as described herein.