Arja Uotila

Human transfer factor in vitro: identification of L-serine and glycine as components with augmenting effect in lymphocyte transformation assay.

The components of human leucocyte dialysate thought to contain transfer factor, which irresponsible for its capacity to augment antigen or mitogen induced lymphocyte blasogenesis in vitro, were determined by comparing the activity of the original dialysate with its fractions derived from chromatography on Sephadex G‐10. Augmentation of leucoagglutinin (LA) induced transformation was only observed using the original dialysate and two fractions (IIb and IIc) eluting before the total volume of the column. These fractions contained the majority of free amino acids in the dialysate, and no trichloroacetic acid (TCA) precipitable material. Synthetic mixtures, containing the same amount of free amino acids as the fractions, augmented both tuberculin PPD and LA induced blastogenesis in a similar manner to the fractions. The augmenting activity of the synthetic mixture and hence, presumably, the dialysate fractions seemed to be due mostly to l‐serine and glycine, non‐essential amino acids not present in MEM‐S, the culture medium used. No augmenting affect by leucocyte dialysate was seen when MEM‐S was replaced by RPMI 1640, a culture medium containing both l‐serine and glycine. The results suggest that the non‐specific in vitro activity of leucocyte dialysate in this test may be due to a medium supplementation effect and that this phenomenon is probably not relevant to the nonspecific consequences of injection of transfer factor in vivo.

The Effect of Transfer Factor on Lymphocyte Transformation. Comparison of Augmentation by Dialysates of Leucocytes and Lymphoid and Non-Lymphoid Organs

In this paper we describe experiments to determine whether dialysable extracts of non-lymphoid and lymphoid organs augment lymphocyte transformation in vitro in a manner similar to the augmenting activity of leucocyte dialysates. Human peripheral blood leucocytes were cultured with tuberculin PPD or leucoagglutinin, and dose-related effects of the dialysable extracts on lymphocyte transformation were studied by 125IUdR incorporation. Augmentation of lymphocyte transformation was obtained not only with leucocyte dialysates but also with dialysable extracts of lymphoid and non-lymphoid organs (e.g. spleen, liver, kidney, brain). It is concluded that the agent or agents present in dialysable leucocyte transfer factor preparations, which augment lymphoid transformation in vitro, are widely distributed throughout mammalian tissues.

Studies on the Chemical Composition and Biologieal Properties of Transfer Factor

Dialyzable transfer factor (dTF) was fractionated on Sephadex G-10 and G-25 fine columns, and biological activity was found in 3 fractions. One of these, designated VIa, and having a tendency to adsorb to the Sephadex G-10 gel, was shown to have a therapeutic effect on certain immunological diseases. Analysis of this fraction on thin-layer and gas chromatography and with infrared and mass spectroscopy indicated that about half of this fraction was composed of uracil; additional unidentified heterocyclic and aromatic substances were present in this fraction. Adjacent fraction V contained tyrosine and a small polyribonucleotide, and fraction VII hypoxanthine and additional unidentified components. Our results suggest that the therapeutic activity of dTF is not mediated through an immunologically specific informational molecule, but is rather based on non-specific stimulation of the expression of the immune response.

Studies on the Biological and Chemical Nature of a Component in Transfer Factor with Immunologically Nonspecific Activity

Publisher Summary This chapter focuses on the studies of the biological and chemical nature of a component in transfer factor with immunologically nonspecific activity. Fractionation of human leucocyte dialysate on Sephadex G-10 yielded seven fractions and fractions I, III and VI induced skin reactivity to oidiomycin in negative recipients. Fraction VI had therapeutic effect in various immune deficiency states, characteristically its skin conversion effect was not associated with increase in the in vitro blastogenic response to antigenic stimulus in the recipients, but low PHA responses as well as low E rosette values normalized occasionally. In order to purify the active component dialyzates were fractionated on three consecutive columns of Sephadex G-10, resulting in two new fractions and several subs fractions. The UV absorbing spot in fraction via had the same RF value as uracil and IB-spectra, mass spectra as well as NMR-FT analysis indicated that uracil was the major, but not only component in this fraction. Adjacent fraction VII contained hypoxanthine, but again this was not the only component present.

Studies on the chemical nature of dialysable transfer factor. Comparison of human leukocyte dialysate and dialysates derived from human serum and from mammalian lymphoid and non-lymphoid organs.

The chemical nature of human dialysable transfer factor (TFd), capable of augmenting delayed hypersensitivity (DH) in human recipients, and some mammalian organ dialysates, known to augment DH in antigen-primed guinea pigs, were compared using chromatography on Sephadex G-10 and G-25 columns and on thin-layer plates. The fractions of human leukocyte dialysate which eluted at or after the Vt of the Sephadex columns have previously been shown to contain the in vivo TFd-activity and therefore special attention was paid to corresponding dialysate fractions. All together 52 identified or unidentified components were found at or close to this elution region with thin-layer chromatography (TLC). The 14 identified substances were nucleobases, nucleosides, sugars and aromatic or heterocyclic amino acids. Unidentified components had similar staining characteristics as the identified ones on TLC. No evidence was found for the presence of peptides or nucleotides. There were no components specific for human leukocyte dialysate. Several of the identified and unidentified substances in fractions of humal dialysable leukocyte extract were common to all or nearly all dialysates. The possibility that some of the unidentified components might be responsible for the in vivo effect of human leukocyte dialysate in man or guinea pig is discussed.

Nomenclature for Factors of the HLA System – 1977

This article gives the decisions of the WHO nomenclature committee on leukocyte antigens, in particular concerning (a) the upgrading of certain HLA-A and HLA-B specificities to full HLA status, (b) the designation of new provisional specificities of the HLA-B, HLA-C, and HLA-D loci, and (c) the establishment of a nomenclature for the new specificities identified by serological techniques on B lymphocytes.

Demonstration of a Substance Chemically Similar to Human Transfer Factor in Lymphoid Cell Dialyzates from Several Animal Species

Publisher Summary This chapter elaborates the demonstration of a substance chemically similar to human transfer factor (TF) in lymphoid cell dialyzates from several animal species. Adsorption chromatography and TLC were used for isolation and further characterization of a substance chemically similar to human transfer factor in dialyzates of human leucocytes, and lymphoid organ homogenates from several animals. In adsorption chromatography, similar elution patterns were obtained from dialyzates of human, canine, bovine, ovine and porcine peripheral blood leucocyte lysates and spleen homogenates. In all of these animals one could demonstrate the presence of a substance which by elution volume, 260/280 ratio, RF values in TLC and by staining reactions was similar to the substance isolated from human leucocyte dialysates, and shown to have TF activity. Infrared, and mass spectrometry were used for further chemical analysis of the isolated substances. The results indicate that the substance is closely related, if not identical with uracil, a finding which is in accordance with those obtained with human leucocyte dialyzates.