Transfer factor: an overlooked potential for the prevention and treatment of infectious diseases.

Transfer factor (TF) is a low-molecular-weight lymphocyte extract capable of transferring antigen-specific cell-mediated immunity (CMI) to T lymphocytes. It has been used successfully as an adjuvant or primary therapy for viral, parasitic, fungal, and some bacterial infections, as well as immunodeficiencies, neoplasias, allergies and autoimmune diseases. From the list of infections that seem to respond noticeably to transfer factor, those due to viruses of the herpes family are particularly remarkable. Indeed, for these viruses it was shown that TF can prevent infection or relapse, acting as a CMI vaccine. Data also suggest its possible use for adjuvant treatment and probably prevention of two currently widespread infections: tuberculosis and AIDS. Furthermore, TF has an interesting potential: answering the challenge from unknown pathogenic agents, a black box effect permitting production of antigen-specific TF to a new pathogen, even before its identification. It thus seems that the preventative potential of transfer factor is as important as its therapeutic one, both discussed in this review.

Quantitative studies of phytohemagglutinin-induced DNA and RNA synthesis in normal and agammaglobulinemic leukocytes.

Leukocytes from nine patients with acquired agammaglobulinemia were studied in vitro. Synthesis of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) induced by phytohemagglutinin was measured by determination of the degree of incorporation of labeled precursor. Synthesis of both DNA and RNA was decreased in the agammaglobulinemic cells. The presence of an inhibitor in the patients' sera could not be demonstrated. These results suggest that the basic defect in agammaglobulinemia is cellular rather than humoral.

Structural studies of human gamma-G-myeloma proteins of different antigenic subgroups and genetic specificities.

1. Peptide maps of Fc fragments or heavy chains of 36 G myeloma proteins and two "heavy chain disease" proteins belonging to the four gamma-chain subgroups revealed very striking similarities between them. However differences in a few peptides were noted. This was most pronounced for the Ge(gamma(2)d) subgroup which lacked three peptides characteristic of the other three subgroups. While Fc fragments from different proteins belonging to the same subgroup appeared very similar, minor differences in addition to those based on currently recognized Gm factors were occasionally noted. 2. Fc fragments from Gm(a+) We(gamma(2)b) proteins had a peptide previously shown to be characteristic of normal Gm(a+) gammaG-globulins. Fc fragments from Gm(a-) molecules belonging to the We(gamma(2)b), Vi(gamma(2)c), or Ne(gamma(2)a) subgroups, whether Gm(b+), Gm(f+), or Gm(-), had the peptide previously identified in Gm(b+f+) normal gammaG-globulin. This "non-a" peptide was absent in peptide maps from Gm(-) molecules of the Ge(gamma(2)d) subgroup which contained instead another peptide with the same electrophoretic mobility but migrating slightly further on chromatography. 3. Both the "a" and "non-a" peptides were pentapeptides having three amino acids in common, and differing in the other two. The "a" peptide contained one residue of lysine, aspartic acid, threonine, leucine, and glutamic acid. The "non-a" peptides prepared from Gm(b+), Gm(f+), and Gm(-) proteins were identical and contained one residue of lysine, threonine, and methionine sulfone, and two residues of glutamic acid. 4. Several possible mechanisms for the origin of these differences, and their possible role in serologic specificity are discussed.

IgM-induced tumor cell cytotoxicity mediated by normal thymocytes.

In summary, we have found that IgM, from mice which have undergone regression of primary MSV tumors, will induce cytotoxicity against the appropriate target cells by normal splenocytes and normal thymocytes. The thymocyte-induced cytotoxicity i

Immunohistological and elution studies of the human placenta.

An immunohistological survey of 28 full-term human placentas has demonstrated deposits of IgG, beta1C, beta1E, and fibrinogen/fibrin in areas of fibrinoid necrosis and on the trophoblast basement membrane in approximately 35% of placental villi. Traces of IgM were detected at similar sites in 18 of 28 full-term placentas. In 11 specimens of immature placentas (10-18 wk gestation) traces of IgG and beta1C and deposits of fibrinogen/fibrin were also present, but IgM was not detected in this material. IgG was recovered in acidic eluates from an homogenized placenta which behaved as an antibody reactive with unidentified material present in fibrinoid deposits and on the thickened trophoblast basement membrane of some villi. It could not be determined whether this IgG was derived from the maternal or fetal circulation.

Lymphocyte culture: induction of colonies by conditioned medium from human lymphoid cell lines.

The presence of phytohemagglutinin or pokeweed mitogen in cultures of human peripheral blood mononuclear cells in agar is known to stimulate the formation of lymphoid colonies. We now report that similar colonies can be induced in the absence of plant lectins upon addition of filtered and ultracentrifuged conditioned medium (CM) obtained from certain human lymphoblastoid cell lines. Colony formation required at least 6 X 10(5) mononuclear cells per milliliter, and optimum results were obtained at concentrations of 1 X 10(6) cells/ml in the presence of 20% CM (50-500 colonies per 10(6) cells cultured). Individual cells within colonies displayed uniform morphological characteristics of lymphoid cells, and the majority formed rosettes with sheep erythrocytes, suggesting that they were of T-cell type.

Heterozygosity at Gm loci associated with humoral immunity to osteosarcoma.

Serum samples from 50 Caucasian patients with osteosarcoma were tested for the presence of antibodies to osteosarcoma-associated antigens (OSAA) and typed for nine Gm markers. A highly significant association was found between Gm 3;5,13,14 and unresponsiveness to OSAA, and between 1,3,17;5,13,14,21 and responsiveness to OSAA. These results suggest the existence of complementary immune response genes which in the heterozygous condition permit a response to OSAA.

Polyclonal activation of human peripheral blood B lymphocytes by formaldehyde-fixed Salmonella paratyphi B. I. Immunoglobulin production without DNA synthesis.

A "new" polyclonal activator of human peripheral blood B cells, formaldehyde-fixed Salmonella paratyphi B, is described. This bacterium does not stimulate cell proliferation as measured by incorporation of tritiated thymidine but does stimulate a subpopulation of B cells to secrete large amounts of IgM, IgG, and IgA in 7-day cell cultures. The immunoglobulins (Ig) produced by cells responding to S. paratyphi B are not specific antibodies against the bacterial antigens. In comparison with other B cell activators (pokeweed mitogen, Staphylococcus aureus Cowan I, and lipopolysaccharide), S. paratyphi B stimulation produced greater amounts of IgM but less IgG than pokeweed mitogen (PWM) or S. aureus Cowan I; lipopolysaccharide failed to stimulate significant Ig production on day 7 in most cases. In addition, the response to S. paratyphi apparently did not require T cell collaboration. These results suggest that the B cell subpopulation(s) responding to S. paratyphi B may be more differentiated B cells than those responding to either PWM or S. aureus Cowan I. Peripheral blood mononuclear cells from five patients with common variable immunodeficiency without evidence of abnormal suppressor T cells or monocytes failed to respond to S. paratyphi B, whereas cells from two of the same patients responded well to S. aureus Cowan I and partially to PWM. Thus, S. paratyphi B appears to be superior to other B cell activators for studies of B cell function in normal and abnormal states.

Peyer's patches: immunologic studies.

The immune capabilities of the Peyer's patches have been investigated by the use of an in vitro system. Despite our failure to stimulate Peyer's patch lymphocytes in vivo it appears that Peyer's patches behave immunologically as peripheral lymphoid tissues. Cultures prepared from the dissociated Peyer's patches of normal rabbits respond to sheep erythrocytes. The response is comparable to that obtained with spleen cultures from the same animals and is not dependent on the presence of the epithelial cells which line the lumen. Similar thymic cultures do not respond. Our experiments with cultures prepared from rabbits which have received one or two injections of SRC show that the Peyer's patches contain both IgM and IgG "memory" cells which have migrated from the spleen. The concentration of these cells in the spleen remains several hundredfold higher.

Studies on the immune response to a characterized antigenic determinant of the tobacco mosaic virus protein.

THE FOLLOWING PEPTIDES HAVE PREVIOUSLY BEEN SHOWN TO BIND SPECIFICALLY WITH ANTIBODIES TO TMVP: (a) An eicosapeptide representing residues 93-112 of TMVP and having the sequence Ileu-Ileu-Glu-Val-Glu-AspNH(2)-GluNH(2)-Ala-AspNH(2)-Pro-Thr-Thr-Ala-Glu-Thr-Leu-Asp-Ala-Thr-Arg. (b) Its C-terminal decapeptide. (c) Its C-terminal pentapeptide. (d) N-octanoyl-C-terminal-tripeptide. (e) (Lys)(4)-C-terminal-pentapeptide. (f) (Lys)(7) C-terminal-pentapeptide. The present communication deals with the investigation of several parameters of the immunological activity of the peptides. The results show that none of the peptides tested were immunogenic in guinea pigs, nor did they stimulate the incorporation of (14)C-thymidine by spleen cells derived from TMVP-primed animals. Results also showed that all of the peptides tested could elicit specific delayed and immediate skin reactions in TMVP-sensitized guinea pigs, and furthermore, that the peptides could specifically inhibit the migration of peritoneal exudate cells derived from these animals. The elicitation of delayed skin reactions and the ability to inhibit migration of peritoneal exudate cells were independent of carrier specificity.

Experimental allergic encephalitis. Dissociation of cellular immunity to brain protein and disease production.

The encephalitogenic determinant of brain protein, a nonapeptide having the amino acid sequence Phe-Ser-Trp-Gly-Ala-Glu-Gly-Gln-Lys, has been characterized and synthesized. In a previous study, analogues of this encephalitogenic peptide were synthesized and some were shown to be encephalitogenic while others were not. Guinea pigs were immunized with encephalitogenic peptides having amino acid sequences different from that in the native protein. These guinea pigs did not show cellular immunity in vivo (skin reactivity) or in vitro (lymphocyte stimulation or macrophage migration inhibition) to the encephalitogenic brain protein (EP) although they did show cellular immunity to the immunizing antigenic peptide. Guinea pigs immunized with an encephalitogenic peptide having the same amino acid sequence as the brain protein, or with a nonencephalitogenic peptide having the same amino acid sequence as the native protein but lacking the terminal lysine, did develop cellular immunity to the EP. Animals immunized with EP showed cellular immunity to this protein, but not to the encephalitogenic peptides. Animals immunized with nonencephalitogenic protein (NEP), prepared by altering the tryptophan residue of EP, did not develop disease but did show cellular immunity in vitro and in vivo to the EP. Animals protected from disease by immunization with NEP similarly showed cellular immunity to EP. Thus, the results suggest a dissociation between cellular immunity to EP and the production of experimental allergic encephalitis (EAE). Animals immunized with the encephalitogenic peptides develop EAE, but do not show cellular immunity to EP, and animals immunized with NEP show cellular immunity to EP but do not develop EAE. A fresh approach to the examination of the pathogenesis of EAE is now possible through the use of these well-characterized antigens.

Mycoplasma inhibition of phytohemagglutinin stimulation of lymphocytes.

Goat lymphocytes were cultured in vitro with phytohemagglutinin and nonviable mycoplasmas. Addition of the mycoplasmas, even as late as 45 hours after adding phytohemagglutinin, completely inhibited the increase in synthesis of DNA and RNA normally induced in lymphocytes by the mitogen. The suppression of synthesis did not result from killing of the cells by the mycoplasmas, combination of the organisms with phytohemagglutinin, or competition for combining sites on the cell surface, which indicates that some other mechanism of inhibition was operative. A similar depression of response to phytohemagglutinin in lymphocytes in culture has been observed in human diseases associated with an immune defect. The present demonstration that at least certain mycoplasmas can profoundly affect lymphocyte function in vitro suggests that thay may alter the immune response in vivo.

Phencyclidine-induced immunodepression.

Phencyclidine ("PCP" or "angel dust") and some of its derivatives are psychotomimetic drugs that have been used in general anesthesia for some time. This drug blocks potassium ion channels in brain tissue, and there is a specific PCP binding to lymphocytes. In a study of the effects of this drug on immunocyte function, it was found that humoral and cellular immune responses in vitro were depressed when immunocytes were treated with PCP before biological assay. This finding has implications for PCP abuse and also for the use of its derivative in general anesthesia, where it may contribute to postoperative infection.

Secretory immunoglobulin A: autoantibody activity in gastric juice.

An immunoglobulin A of the secretory variety, present in the gastric juice of a patient with pernicious anemia, was shown to have specificity for intrinsic factor. This is the first demonstration in gastric juice of antibody activity restricted to secretory IgA; further, this is the first example of an exocrine (gastric) immune system producing an autoantibody specifically directed toward a product synthesized by that same exocrine organ.

Preferential synthesis of the G1m(1) allotype of IgG1 in the central nervous system of multiple sclerosis patients.

Quantitations of the G1m(1) and G1m(3) allotypic determinants of human immunoglobulin G were performed by radioimmunoassay on cerebrospinal fluid and serum samples from patients with multiple sclerosis and from patients with other neurological disorders. In multiple sclerosis patients that were heterozygous for these determinants, G1m(1) concentration in the cerebrospinal fluid was greatly increased-reflected by an increased ratio of G1m(1)-in comparison with that of patients with other neurological disorders. These results suggest that in the heterozygous multiple sclerosis patients, most of the plasma cells in the central nervous system that secrete oligoclonal immunoglobulin G preferentially synthesize G1m(1) IgG1 molecules.

Factor 8 detection by hemagglutination inhibition: hemophilia A and von Willebrand's disease.

Factor VIII activity was detected immunologically in both the serums and plasmas of 14 normal individuals and 14 patients with hemophilia A. A hemagglutination-inhibition test with rabbit antibody to highly purified (10,000-fold) factor VIII from humans was used. Serums and plasmas from eight patients with von Willebrand's disease showed little or no factor VIII activity in this test, an indication that the test may serve as a specific assay for differentiation between von Willebrand's disease and hemophilia A.

Human monocytes: distinct receptor sites for the third component of complement and for immunoglobulin G.

Human monocytes contain two distinct receptor sites, one specific for the third component of complement (C'3), the other for immunoglobulin G(gammaG). The two receptors may function either independently or cooperatively in the induction of phagocytosis. Ingestion of erythrocytes coated with immunoglobulin M antibody requires a relatively large number of bound C'3 molecules per cell. Ingestion of erythrocytes sensitized with gammaG antibody is independent of complement; however, the reaction is inhibited by concentrations of gammaG far below those in normal serum. Inhibition by gammaG-globulin is overcome by a relatively small number of bound C'3 molecules per cell. The two monocyte receptors exert a cooperative effect on ingestion by monocytes of erythrocytes coated with gammaG antibody in the presence of inhibitory amounts of free gammaG.

Thymus-derived rosette-forming cells in various human disease states: cancer, lymphoma, bacterial and viral infections, and other diseases.

Lymphocytes that bind in vitro to sheep erythrocytes in a rosette formation are thymus-derived. A modified technique that does not detect the total number of rosette-forming cells (RFC) was used to study normal subjects and various disease states. Of 100 healthy subjects, 95 had more than 15% RFC (mean 28.4+/-6.5%). We studied 104 patients with solid tumors, who were classified according to clinical status and stage of therapy. Of 19 newly diagnosed patients, 13 had less than 15% RFC. Of 44 untreated patients undergoing relapse, 32 had less than 15% RFC. In both categories, patients with metastases had fewer RFC than patients with localized disease. 11 patients were studied 2 wk after cessation of therapy; four of them showed less than 15% RFC. Only one of 30 patients in remission had less than 15% RFC. In seven patients followed for various periods of time, the numbers of RFC correlated generally with clinical status. 11 patients with chronic lymphatic leukemia had very low percentages of RFC. 21 of 21 patients with symptoms of viral upper respiratory diseases had less than 15% RFC. RFC returned to normal values between 5 days and 7 wk after disappearance of clinical symptoms. 20 patients with bacterial infections had normal numbers of RFC. Of 25 patients with miscellaneous nonimmunologically related diseases, two had low numbers of RFC. It appears that the percentage of RFC may be valuable in evaluating not only immunological defenses but also the status of patients with solid tumors, lymphomas, viral diseases and, perhaps, bacterial infections.

Restoration of immune responses of aging hamsters by treatment with isoprinosine.

Immune competence declines with advanced age in hamsters, as in other laboratory mammals and in humans. We found significant alterations in the functional parameters of different populations of immunocytes (natural killer cells, T cells, monocytes, and suppressor cells) in aging hamsters, beginning at approximately 14 mo of age. Natural killer cytotoxicity, phytohemagglutinin-induced lymphocyte stimulation, and monocyte chemotaxis were decreased in aging Lak:LvG(Syr) outbred hamsters. When old hamsters were given a single injection (5 mg/kg body wt) of isoprinosine, a chemical immune potentiator, these three immune parameters increased almost to the levels found in young adult hamsters but returned to pretreatment levels after 7 d. Suppressor cell activity for the lymphocyte response to phytohemagglutinin, which increased with age, was decreased after treatment. In old hamsters treated with weekly injections of isoprinosine, these four immunological parameters remained at or near the levels found in young adults.

The human rosette-forming cell as a marker of a population of thymus-derived cells.

Sheep red blood cells can surround, in vitro, some human peripheral blood lymphocytes in a formation called a rosette. The number of rosetteforming cells (RFC) in 50 normal persons had a wide range (4-40%). The organs of 13 human fetuses (11-19 wk conceptional age) were examined for the presence of RFC. The thymus possessed the highest percentage of RFC, the maximum being 65% of total thymocytes in two 15-16 wk fetal specimens. Blood RFC were always present and their number slightly increased in the oldest fetuses. The bone-marrow showed 0-8% in the six fetuses studied. RFC were found in the spleen around the 13th wk and in the liver around the 17th wk of gestation. These observations lead to the hypothesis that human blood RFC may be chiefly thymic derived. Studies of patients with immunological disorders support this hypothesis: one patient with Nezelof syndrome had no blood RFC and four patients with Wiskott-Aldrich syndrome had a low number of blood RFC (1 and 1.5%). Patients with acquired hypogammaglobulinemia showed a normal percentage of RFC. With the fetal thymocytes, the percentage of inhibition with anti-mu serum increased with the fetal age to become complete in the oldest fetuses studied. Incubation of the oldest fetal thymocytes or the blood lymphocytes with anti-gamma serum of anti-mu serum completely inhibited the rosette formation. These results suggest that mu-chain determinants are present on human fetal thymocytes and blood RFC. The significance of the presence of gamma-chain determinants on these cells is unclear.