Transfer Factor: Myths and Facts.

Transfer factor (TF), also called "Lawrence transfer factor", or dialyzable leukocyte extract (DLE), has been used since the mid-twentieth century to transfer specific skin hypersensitivity through the injection of leukocytes from immunized donors to animals and humans. The main mechanism of action of TF has been suggested at the level of cell-mediated immunity, as it induces the production of migration inhibitory factor (MIF) and interferon gamma (IFN-γ). Otherwise, TF can inhibit nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB), and decrease tumoral necrosis factor α (TNF-α) and IL-4 levels. Given these biological effects, TF has been prescribed for a wide variety of conditions including infections, allergies, autoimmunity, and cancer, with inconsistent results. The exact nature of TF, however, remains unknown, so it has been impossible to accurately define its pharmacokinetics or dosage. This is further complicated because researchers have used TF in a variety of ways across the different studies: antigen-specific or non-antigen-specific, orally or subcutaneously administered, human and non-human origin. In this review we summarize the most important data about what TF is, its mechanism of action, how it is produced, its biological effects, and the available clinical trials using it, in order to establish its role and potential clinical applications in modern medicine.

Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®).

Transferon® is an immunomodulator made of a complex mixture of peptides from human dialyzable leucocyte extracts (hDLEs). Development of surrogate antibodies directed to hDLE is an indispensable tool for studies during process control and preclinical trials. These antibodies are fundamental for different analytical approaches, such as identity test and drug quantitation, as well as to characterize its pharmacokinetic and mechanisms of action. A previous murine study showed the inability of the peptides of Transferon® to induce antibody production by themselves; therefore, in this work, two approaches were tested to increase its immunogenicity: chemical conjugation of the peptides of Transferon® to carrier proteins and the use of a rabbit model. Bioconjugates were generated with Keyhole Limpet Hemocyanin (KLH) or Bovine Serum Albumin (BSA) through maleimide-activated carrier proteins. BALB/c mice and New Zealand rabbits were immunized with Transferon® conjugated to KLH or nonconjugated Transferon®. Animals that were immunized with conjugated Transferon® showed significant production of antibodies as evinced by the recognition of Transferon®-BSA conjugate in ELISA assays. Moreover, rabbits showed higher antibody titers when compared with mice. Neither mouse nor rabbits developed antibodies when immunized with nonconjugated Transferon®. Interestingly, rabbit antibodies were able to partially block IL-2 production in Jurkat cells after costimulation with Transferon®. In conclusion, it is feasible to elicit specific and functional antibodies anti-hDLE with different potential uses during the life cycle of the product.

Lawrence Transfer Factor: Transference of Specific Immune Memory by Dialyzable Leukocyte Extract from a CD8+ T Cell Line.

Lawrence transfer factor (TF) is defined as dialyzable leukocyte extract (DLE) that can transfer antigen-specific cell-mediated immunity from a person testing positive for the antigen in a delayed type hypersensitivity skin test manner to a person negative for the same antigen. A recent article by Myles et al1 has identified a DLE isolated from an established CD8+ T cell line capable of transferring antigen-specific immunity. The DLE contains a portion of the beta chain of the T cell receptor and additional nucleotide and protein factors that are being subjected to further modern biochemical analysis. After months of study that included interviews of TF physician-scientists, we conclude that an antigen-specific TF exists for most, if not all, antigens. By working from a CD8+ T cell line with modern biochemical technology, it should be possible to identify and patent products capable of treating infectious diseases, antigen-responsive cancers, and autoimmune disorders.

Preparation and identification of transfer factor specific to Staphylococcus aureus in vitro.

The objective of the study was to explore the methods for preparing transfer factor specific to Staphylococcus aureus (SA-STF) in vitro. Under the optimum conditions, the spleen cells of rabbits were immunized with SA in vitro to prepare SA-STF, and the immune activities were identified with the phagocytosis and sterilization, skin delayed-type hypersensitivity, and immune protection tests. The concentration of polypeptide was 2.26 ± 0.27 mg/mL, and ribose was 0.684 ± 0.094 mg/mL. The phagocytosis and sterilization rates of the STF group were 70.9 ± 12.4% and 62.1 ± 12.2%, respectively, and compared with the non-specific transfer factor (NTF) group, there were no significant differences (P = 0.074 and 0.069, respectively). The skin was inflamed and marked nodules formed at the injection site in the mice of the STF group rather than the NTF and control groups. The survival rate of the STF-1 group was significantly higher than the survival rates of the STF-2 (P = 0.024) and NTF groups (P = 0.016). SA-STF was prepared and characterized successfully in vitro, and it probably is a biological candidate for therapy or adjuvant therapy for diseases caused by Staphylococcus aureus.

Preparation and properties of the specific anti-influenza virus transfer factor.

Specific anti-influenza virus and normal transfer factors prepared in an experimental animal model, the pig, have been tested for their components, characteristics, and activity of known specificity. Two transfer factors are small molecular mixture which consist entirely or partly of polypeptides and polynucleosides. Moreover, the biological activity of transfer factors could be approved by Rosettes test and specific skin test. The study would lay a foundation for the research and development of other specific transfer factor.

[Transfer factor specific to mouse lung carcinoma of Lewis].

Factor of transfer was identified and sampled in the course of immunization of rats with cells or tissues of mouse lung carcinoma of Lewis and its antitumor action investigated on models of passive and spontaneous dissemination in mice C57BL/6. Samples obtained at the peak of immunological response (day 14) and immunological memory inception (day 60) were shown to be capable of marked antimetastatic effects: fall in metastatic frequency, number and size of metastases correlated with higher index of metastasis formation.

[Transfer factor of immune reactivity to diphtheria-tetanus anatoxin modulates the action of neurotransmitters in the intestinal smooth muscle].

Transfer factor (TF) of immune reactivity (10(-5) - 10(-3) mg/ml) to diphtheria-tetanus anatoxin modulates slow waves and spontaneous contractile activity of non-atropinized smooth muscle stripes (SMS) of guinea-pig taenia coli. TF (10(-4) mg/ml) transforms slow waves into stable depolarization and tonic contraction. After SMS atropinization, the substance acts in the same way. In the presence of methylene blue (10(-5) M), a guanylatecyclase blocker, FT induces transitory increase of SMS muscle tone, which is followed by their stable relaxation. ATP and UTP, purinoceptors agonists, evoke substantial hyperpolarization of smooth muscle cells membrane and their relaxation. FT enhances post-inhibitory excitation in SMS. In the presence of acetylcholine (10(-5) M) FT (10(-4) mg/ml) transforms the inhibitory ATP action on tonic contraction into excitative. This substance (10(-5), 10(-4) mg/ml) enhances Ca2+ mobilization from ryanodine-sensitive calcium store, inhibits the release of these cations from IP3-sensitive calcium store of sarcoplasmic reticulum. TF demolishes the inhibitory actions of sodium nitroprusside (nitric oxide donor), and noradrenaline in taenia coli smooth muscles.

Preparation and determination of immunological activities of anti-HBV egg yolk extraction.

To prepare an effective immune preparation to treat hepatitis B, hens were immunized with hepatitis B vaccines, and then anti-HBV egg yolk extraction (anti-HBV EYE) was refined from egg yolk by a dialyzable method. Its chemical characteristics were identified by ultraviolet spectrum, HPLC, Lowry analysis and pharmacopocia-raleted methods. The specific immunological activity was examined by leukocyte adherence inhibition (LAI) in vitro and delayed type hypersensitivity (DTH) in vivo. Anti-HBV EYE was a small dialyzable substance with molecular weight less than 12 kD containing 18 kinds of amino acids. The preparation could obviously inhibit LAI and DTH which was similar to hepatitis B virus-specific transfer factor of pig spleen. However, there were no similar effects observed in the nonspecific transfer factor (NTF) group, control egg yolk extraction (CEYE) group and hepatitis A virus (HAV) group. The results suggested that anti-HBV EYE contained hepatitis B virus-specific transfer factor (STF) and had the antigen-specific cell immune activity similar to PSHBV-TF. The STF obtained from egg yolk of the hens immunized with specific antigen, might be a potential candidate for immunoregulation in hepatitis B prevention and treatment.

Transfer factors as immunotherapy and supplement of chemotherapy in experimental pulmonary tuberculosis.

Problems of logistics, compliance and drug resistance point to an urgent need for immunotherapeutic strategies capable of shortening the current six month antibiotic regimens used to treat tuberculosis. One potential immunotherapeutic agent is transfer factors. Transfer factors (TF) are low molecular weight dialysable products from immune cells which transmit the ability to express delayed-type hypersensitivity (DTH) and cell mediated immunity from sensitized donors to nonimmune recipients. In this study we determined the efficiency of TF as immunotherapy to treat experimental tuberculosis. When BALB/c mice are infected via the trachea with Mycobacterium tuberculosis H37Rv there is an initial phase of partial resistance dominated by Th-1 type cytokines plus tumour necrosis factor-alpha (TNFalpha) and the inducible isoform of nitric oxide synthase (iNOS), followed by a phase of progressive disease characterized by increasing expression of IL-4, diminished expression of TNFalpha and iNOS, and low DTH. Animals in this late progressive phase of the disease (day 60) were treated with different doses of TF (one injection per week) obtained from spleen cells when the peak of immune protection in this animal model is reached (day 21), or with different doses of TF from peripheral leucocytes of PPD + healthy subjects. We show here that the treatment with murine or human TF restored the expression of Th-1 cytokines, TNFalpha and iNOS provoking inhibition of bacterial proliferation and significant increase of DTH and survival. This beneficial effect was dose dependent. Interestingly, murine TF in combination with conventional chemotherapy had a synergistic effect producing significant faster elimination of lung bacteria loads than chemotherapy alone.

[Analysis of the contents of Zn, Cu, Fe, Mn, Co and Ni in thymopeptide and transfer factor biological injections].

Thymopeptide and transfer factor are two common clinic biological preparations, which are used to cure immunodeficiency, low immunofunction or infectious disease caused by virus or fungi owing to their functions of increasing body immunity. In order to discuss the relationship between trace elements and those related diseases, to coordinate sound clinic use of the preparations and to provide productive data on them, atomic absorption spectrophotometry was used to detect the contents of Zn, Cu, Fe, Mn, Co and Ni in thymopeptide and transfer factor biological preparations. Respective contents of detected elements were compared in a statistical way. Results show that Zn and Co contents in thymopeptide were lower than in transfer factor; the contents of Cu, Fe, Mn and Ni, in thymopeptide were higher than in transfer factor. For Zn, Cu, Mn and Co, there was a significant difference between the two biologicam preparations (P<0.01). For Fe and Ni there was a great difference (Q<0.05). The results can provide useful data for sound clinic biological injection to promote immune function, and for increasing or decreasing certain trace elements in preparation.

Immunological treatments for autism.

Several investigators, including ourselves, have reported significant changes in various immune responses in children with autism. These changes demonstrate dysregulation of the immune system (deficiency in some components of the immune system and excesses in others). In addition, certain genes in the major histocompatibility complex (that regulates immune responses) appear to be involved in autism. Based upon immunological abnormalities, various treatment modalities have been applied to children with autism. In this brief review, these immunological changes and various biological therapies are analyzed and summarized.

Transfer factors: identification of conserved sequences in transfer factor molecules.

BACKGROUND: Transfer factors are small proteins that "transfer" the ability to express cell-mediated immunity from immune donors to non-immune recipients. We developed a process for purifying specific transfer factors to apparent homogeneity. This allowed us to separate individual transfer factors from mixtures containing several transfer factors and to demonstrate the antigen-specificity of transfer factors. Transfer factors have been shown to be an effective means for correction of deficient cellular immunity in patients with opportunistic infections, such as candidiasis or recurrent Herpes simplex and to provide prophylactic immunity against varicella-zoster in patients with acute leukemia. MATERIALS AND METHODS: Transfer factors of bovine and murine origin were purified by affinity chromatography and high performance liquid chromatography. Cyanogen bromide digests were sequenced. The properties of an apparently conserved sequence on expression of delayed-type hypersensitivity by transfer factor recipients were assessed. RESULTS: A novel amino acid sequence, LLYAQDL/VEDN, was identified in each of seven transfer factor preparations. These peptides would not transfer expression of delayed-type hypersensitivity to recipients, which indicates that they are not sufficient for expression of the specificity or immunological properties of native transfer factors. However, administration of the peptides to recipients of native transfer factors blocked expression of delayed-type hypersensitivity by the recipients. The peptides were not immunosuppressive. CONCLUSIONS: These findings suggest that the peptides may represent the portion of transfer factors that binds to the "target cells" for transfer factors. Identification of these cells will be helpful in defining the mechanisms of action of transfer factors.

[The adjuvant and specific activity of transfer factors to Candida albicans antigens]. / Ad'iuvantna ta spetsyfichna aktyvnist' Faktoriv perenosu do antyheniv Candida albicans.

Activity of transfer-factor (TF) delayed type hypersensitivity in allogenic and xenogeneic have studied. Guinea pigs was been sensibilized Soluble antigens and whole cells Candida albicans for production TF. Both TF expressed non specific and specific activity in the guinea pig and mouse leucocytes inhibition migration in vitro and delayed type hypersensitivity in mica. Activity of TF to cellular antigen was lower then TF to soluble antigen in xenogeneic systems. However TF activity was some low for whole cells in xenogeneic systems.

Transfer factor.

AIDS: Transfer factor, a natural substance of the immune system, was discovered in 1949. More than 3,000 scientific articles have established it as an effective treatment for many diseases, usually those related to the immune system. In China, more than six million people have used transfer factor as a prophylaxis for hepatitis. Information on ordering articles on transfer factor, olive leaf extract, and coconut oil is included.^ieng

[Human specific transfer factor to Staphylococcus aureus antigens]. / Liuds'kyi spetsyfichnyi Faktor perenosu do antyheniv Staphylococcus aureus.

Immunobiological properties of human specific transfer factor (TF) to Staphylococcus aureus antigens were studied. It is shown that this TF activated human leucocytes in vitro as well as in vivo. Antigen specificity of TF's immunomodulating effects is also shown. In vitro we used leucocyte migration inhibition test (IML), macrophage inhibition test (MPI) and rosette formation (E-ros). For testing in vivo we used delayed type hypersensitivity (DTH) skin tests.

Transfer factor--current status and future prospects.

We have detected new clues to the composition and function of "Transfer Factor" using the direct Leucocyte Migration Inhibition (LMI) test as an in vitro assay of Dialysates of Leucocyte Extracts (DLE). This approach has revealed two opposing antigen-specific activities to be present in the same > 3500 < 12,000 DA dialysis fraction - one activity is possessed of Inducer/Helper function (Inducer Factor). The opposing activity is possessed of Suppressor function (Suppressor Factor). When non-immune leucocyte populations are cultured with Inducer Factor they acquire the capacity to respond to specific antigen and inhibition of migration occurs. This conversion to reactivity is antigen-specific and dose-dependent. When immune leucocyte populations are cultured with Suppressor Factor their response to specific antigen is blocked and Inhibition of Migration is prevented.

Transfer factor in the age of molecular biology: a review.

Current data suggests that the transferring of immunologically specific information by transfer factor molecules requires interaction with a cell that has been genetically programmed to be antigen reactive but at the time of interaction is unprimed. Contact with transfer factor molecules would allow a naive recipient, on a first encounter with antigen, to make a secondary rather than a primary immunological response. Transfer factor molecules for each and every antigenic determinant are thus necessary. Transfer factors made from animals or humans are capable of transferring antigen specificity across a species barrier. Even primitive species have cells from which one can make transfer factors. The molecules are, therefore, well conserved and it is reasonable to suggest that they are important for normal immunological functioning. Proposed mechanisms of action must explain the fact that transfer factors obtained from the cells of high responder animals are capable of transferring delayed hypersensitivity to low responder animals while the reverse is not true. Transfer factor molecules are likely to interact with the variable regions of the alpha and/or beta chain of T cell receptors to change their avidity and affinity for antigen in a way that otherwise would only occur after an encounter with antigen.

Preliminary observations using HIV-specific transfer factor in AIDS.

Twenty five HIV-1-infected patients, at various stages (CDC II, III and IV) were treated orally with HIV-1-specific transfer factor (TF) for periods varying from 60 to 1870 days. All patients were receiving antiviral treatments in association with TF. The number of lymphocytes, CD4 and CD8 subsets were followed and showed no statistically significant variations. In 11/25 patients the number of lymphocytes increased, whilst in 11/25 decreased; similarly an increase of the CD4 lymphocytes was observed in 11/25 patients and of the CD8 lymphocytes in 15/25. Clinical improvement or a stabilized clinical condition was noticed in 20/25 patients, whilst a deterioration was seen in 5/25. In 12/14 anergic patients, daily TF administration restored delayed type hypersensitivity to recall antigens within 60 days. These preliminary observations suggest that oral HIV-specific TF administration, in association with antiviral drugs, is well tolerated and seems beneficial to AIDS patients, thus warranting further investigation.

Efficacy of transfer factor in treating patients with recurrent ocular herpes infections.

Recurrent ocular herpes is an insoluble problem for the clinician. As cellular immunity plays an important role in controlling herpes relapses, and other studies have shown the efficacy of HSV-specific transfer factor (TF) for the treatment of herpes patients, an open clinical trial was undertaken in 134 patients (71 keratitis, 29 kerato-uveitis, 34 uveitis) suffering from recurrent ocular herpetic infections. The mean duration of the treatment was 358 days, and the entire follow-up period 189,121 before, and 64,062 days after TF treatment. The cell-mediated immune response to the viral antigens, evaluated by the lymphocyte stimulation test (LST) and the leucocyte migration test (LMT) (P < 0.001), was significantly increased by the TF treatment. The total number of relapses was decreased significantly during/after TF treatment, dropping from 832 before, to 89 after treatment, whereas the cumulative relapse index (RI) dropped, during the same period, from 13.2 to 4.17 (P < 0.0001). No side effects were observed. It is concluded that patients with relapsing ocular herpes can benefit from treatment with HSV-specific TF.

Orally administered HSV-specific transfer factor (TF) prevents genital or labial herpes relapses.

Forty-four patients suffering from genital (22) and labial (22) herpes were orally treated with HSV-1/2-specific transfer factor (TF). TF was obtained by in vitro replication of a HSV-1/2-specific bovine dialysable lymphocyte extract. Treatment was administered bi-weekly the first 2 weeks, and then weekly for 6 months, most patients received 2-3 courses. The total observation period for all patients before treatment was 26,660 days, with 544 relapses, and a relapse index of 61.2, whereas the cumulative observation period during and after treatment was 16,945 days, with a total of 121 relapsing episodes and a cumulative RI of 21.4 (P < 0.0001). Results were equally significant when the 2 groups of patients (labial and genital) were considered separately. These observations confirm previous results obtained with bovine HSV-specific TF, and warrant further studies to establish HSV-specific TF as a choice of treatment for preventing herpes recurrences.