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.

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.

[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.

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.

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.

Clinical and immunologic response to antigen-specific transfer factor in drug-resistant infection with Mycobacterium xenopi.

The administration of transfer factor obtained from three donors who had recovered from clinical infections with Mycobacterium xenopi to a patient who had a destructive pulmonary infection with this organism, was associated with the reversal of an unfavorable clinical course. Cavitary tuberculosis associated with resistance to all combinations of antituberculosis drugs was probably related to a concurrent depression of cell-mediated immunity of unknown origin. Antigen specific but not nonspecific transfer factor caused a rapid and prolonged improvement in both the pulmonary disease and the immunologic deficiency. Cross-reactivity between the antigenic determinants of M. xenopi and Mycobacterium tuberculosis made it possible to use transfer factor obtained from donors responsive to purified protein derivative of tuberculin. This study clearly demonstrates the additional benefits to be gained from using transfer factor that is antigen-specific in the treatment of infectious diseases.

Structural nature and functions of transfer factors.

Transfer factors are molecules that "educate" recipients to express cell-mediated immunity. This effect is antigen-specific. The most consistent effects of transfer factors on the immune system are expression of delayed-type hypersensitivity and production of lymphokines such as macrophage migration inhibitory factor (MIF), which is probably identical to gamma-interferon in response to exposure to antigen. Transfer factors bind to antigens in an immunologically specific manner. This discovery has enabled us to isolate individual transfer factors from mixtures that contain several transfer factors. This reactivity probably explains the specificity of individual transfer factors, and it has provided a method for purification of individual transfer factors to apparent homogeneity. The purified materials are immunologically active and antigen-specific. They have molecular weights of approximately 5,000 Da and appear to be composed entirely of amino acids. Transfer factors appear to offer a novel means of molecular immunotherapy for certain patients with defective cell-mediated immunity.

Development of immunological responsiveness and resistance to infection with Treponema pallidum in rabbits given immune lymphocyte preparations.

Three groups of normal rabbits were administered: 1. viable lymphocytic cells; 2. lysates of lymphocytes obtained by freezing and thawing, and 3. low molecular weight dialyzable transfer factor (TF). The lymphocytes were derived from peripheral blood, lymph nodes, and spleens of rabbits: 1. cured of longlasting syphilitic infection by penicillin treatment, and 2. immunized with nonviable T. pallidum, exhibiting a good reactivity by tests for humoral and cell-mediated responses. All the three lymphocyte preparations were found to be able to transfer cell-mediated response as measured by the Macrophage Migration Inhibition Test (MMI) and the Skin Test for Delayed Hypersenitivity to Treponemal Antigens (SDH), from a reactive donor to the nonreactive recipient rabbit; the intensity of the response found in the recipient was roughly the same as in the donor. None of the lymphocyte preparations transferred humoral response, as measured by the TPI and VDRL tests. Only whole lymphocytic cells were found to be able to confer a stat of resistance to infection with T. pallidum on normal recipients.

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 in vitro of tubercolin hypersensitivity by sensitised lymphocytes.

An account is given of the experimental in vitro transfer of PPD-specific delayed hypersensitivity to peripheral leukocytes pulsed with a very small portion of autologous lymphocytes pre-sensitised with PPD-specific transfer factor (TF). Transfer was assessed by means of the leukocyte migration inhibition test, the lymphocyte locomotion test, and the leukocyte adherence inhibition test. These tests were positivized in all the experiments. It is suggested that these results indicate that TF does not act in itself, but triggers a reaction that expands the effect of hypersensitivity.

Immunopotentiation against internal parasites.

The economic importance of livestock losses attributed to internal parasites has led to the need of developing and successfully employing effective antiparasitic vaccines. An increasingly important strategy in the development of more effective vaccines is the preparation and use of immunopotentiators or adjuvants. Immunopotentiators or adjuvants have been used effectively in combination with antigen(s) or alone to protect animals against internal parasites. The use of immunopotentiators for specific and nonspecific stimulation of protective immunity against internal parasites is reviewed.

Successful passive transfer of resistance to Fasciola hepatica infection in rats by immune serum and transfer factor.

Resistance to Fasciola hepatica was shown by a 64 per cent reduction in fluke recovery compared with control rats was recorded in rats inoculated with serum from sheep experimentally infected with F hepatica. A comparable reduction (55.6 per cent) was obtained using transfer factor prepared from lysed leucocytes of infected rats. No significant reduction was obtained using transfer factor prepared from sheep or calves.

Investigations of 'transfer factor' activity in the transfer of immunity to Trichostrongylus axei infections in sheep.

Three investigations are described in which non-dialysed and dialysed leucocyte lysates, 'transfer factor' prepared from the blood of sheep infected with Trichostrongylus axei successfully transferred immunity to challenge infection with that parasite in susceptible lambs. Similar leucocyte lysates from parasite-free lambs failed to transfer a similar resistance to challenge infection. 'Transfer factor' treatment produced a 30--72 per cent reduction in a total worm burden compared to susceptible control lambs. In the first two investigations the donor and recipient lambs were genetically dissimilar and in the third investigation were of different breeds. The resistance transfer was considered to operate independently of immune incompetence.

Investigation of Haemonchus contortus infections in sheep. Comparison of irradiated larvae and transfer factor treatment.

A comparison is made between the resistance conferred to Haemonchus contortus challenge by irradiated larval treatment and transfer factor (TF) treatment in four-month-old lambs and seven-month-old lambs. As in previous investigations the irradiated larval treatment failed to confer resistance to the four-month-old lambs challenged with 10,000 third stage larvae while similar irradiated larval treatment in seven-month-old lambs reduced the worm burdens by 40 per cent compared to controls. The TF treatment produced a 34 per cent reduction in the challenge infection in the four-month-old lambs and a 45 per cent reduction in the seven-month-old lambs compared to the control lambs. It is concluded that TF activity operates independently of immune competence.

Adoptive transfer of delayed wattle reactivity in chickens with a dialyzable leukocyte extract containing transfer factor.

Delayed wattle reactions (DWR) to tuberculin, diphtheria toxoid (DT), and keyhole limpet hemocyanin (KLH) were transferred to chickens with dialyzable leukocyte extracts (DLE) prepared from splenic leukocytes of chickens sensitized and reactive by DWR to the tuberculin, DT, or KLH. A DLE prepared from chickens unsensitized and unreactive to tuberculin by DWR failed to transfer DWR to tuberculin in recipients. Only chickens injected with DLE from tuberculin sensitized and reactive chickens exhibited significant (P less than .01) DWR to tuberculin. Chickens that received DLE prepared from DT sensitized and reactive chickens exhibited significant (P less than .01) DWR to DT but not to KLH. Further, DWR to both DT and KLH was transferred with DLE prepared from chickens sensitized to both antigens. Adoptive transfer of DWR to KLH in comparison to DT was more successful. Finally, the serial transfer of DWR to KLH (P less than .05) but not to DT (P greater than .05) was accomplished using DLE prepared from chickens that previously were recipients of DLE prepared from chickens sensitized to KLH and DT. Results indicate that DLE prepared from chickens contain transfer factor (TF) responsible for adoptive transfer of DWR to tuberculin, DT, and KLH.

Adoptive transfer of delayed hypersensitivity and protective immunity to Eimeria tenella with chicken-derived transfer factor.

Delayed hypersensitivity (DH) and protective immunity were transferred to nonimmune 4- and 10-week-old broiler chickens with transfer factor (TF) prepared from splenic leukocytes of chickens immunized with CocciVac D. Only chickens injected with the immune TF showed DH by wattle reaction to oocyst antigen and protective immunity to Eimeria tenella challenge infection. Chickens given a single injection of TF 5 days before challenge infection exhibited a significant (P less than .05) DH response at the time of infection. Immune TF preparations were active at concentrations of 100, 200, or 400 mg. Neither DH nor protective immunity was transferred to chickens injected with the TF-diluent control or nonimmune TF. The nonimmune TF was prepared from chickens kept free of coccidial infection. These findings indicated that TF prophylaxis produced beneficial results in nonimmune chickens by conferring some protection against challenge with E. tenella. The effects of TF on T-lymphocyte mediated protective immunity to coccidia in chickens are discussed.

Transfer factor therapy: clinical experience and the role of the E rosette assay.

Transfer factor has been administered to 17 patients, most with infectious diseases of various kinds. In 12 patients the therapy was followed by a definite clinical improvement although in most cases conventional chemotherapy was given concomitantly. In all cases where clinical improvement followed the sheep red cell or E rosette assay showed low values initially, with an improvement following therapy. This test of T lymphocyte function may be useful both in predicting patients likely to respond to transfer factor, and in monitoring response to treatment. As no specific assay of transfer factor activity is available, an in vivo rise in E rosette formation following transfer factor administration serves as a crude indicator that the injected material has some biological activity.

Transfer with dialysable transfer factor of T-lymphocyte cytolytic response to tick-bone encephalitis virus antigen in naive mice.

Transfer factor activity was demonstrated in the dialysable extract from lymphocytes from outbred donor mice, in which immunization with live attenuated Langat (E5 "14") virus induced a stage of high resistance against challenge with virulent tick-borne encephalitis virus. Administered to naive inbred recipient mice, the extract conveyed in them specific cytolytic activity, exerted by their T lymphocytes, as demonstrated by the 51Cr release assay on tick-borne encephalitis virus-infected syngeneic L-929 cells.

Nature and antigen-specific activities of transfer factor against herpes simplex virus type 1.

Transfer factor specific for herpes simplex virus (HSV) type 1 (TFHSV-1) was prepared from splenic cells of HSV-1 immunized mice. Protection was transferred with TFHSV-1 to nonimmune mouse recipients. The TFHSV-1 injected mice had a higher survival rate after lethal HSV-1 challenge as compared to mice injected with a nonspecific transfer factor (P less than 0.05). 51Cr-labelled leukocyte adherence inhibition (51-Cr-LAI) test was used to demonstrate the specific activity of transfer factor in vitro. Only leukocytes incubated with TFHSV-1 exhibited significant adherence inhibition (P less than 0.01) to HSV-1 antigen, but not to control antigen. Specific activity component of TFHSV-1 (STFc) was separated by affinity adsorption with the antigen. Activity of STFc in 51Cr-LAI test was significantly higher than that of TFHSV-1 (P less than 0.01). Ratio activity of STFc in protective host immunity was 16 times as much as that of TFHSV-1. STFc was analysed by high performance liquid chromatography, thin layer chromatography and isoelectric focusing in the polyacrylamide gel. Results revealed that STFc appeared to be a polypeptide with a molecular weight of about 12,870 dalton.