Isolation and purification of HSV-1 specific transfer factor produced by HSV-1 immunized goat leukocyte dialysate.

A herpes simplex virus type 1 (HSV-1)-specific transfer factor (TF), was separated and purified from the leukocyte dialysate of goats immunized with HSV-1 using affinity chromatography on antigen-sorbent and reversed phase high performance liquid chromatography (RP-HPLC). The antigen-specific activities of the starting dialysate and the isolated TF component (s) were examined by 51Cr-labelled leukocyte adherence inhibition (51Cr LAI) assay. The analytical hydrophobic interaction HPLC (HI-HPLC) and isoelectric focusing (IEF) techniques were employed to evaluate the purity and the isoelectric point (PI) of isolated TF component(s). The experiments provided a two-step procedure for purifying the TF material from the starting dialysate. It seems that the purified active TF component (PTFC) was specific for HSV-1. The specific PTFC activity was increased 10,000-fold as compared with the activity of the dialysate. The active moiety appeared as a single band in the IEF gel as demonstrated by silver staining; it was hydrophilic and its PI was pH 4.48.

Chemical characterization of the purified component of specific transfer factor in the leukocyte dialysates from HSV-1 immunized goats.

The chemical characterization of the purified component responsible for HSV-1 specific transfer factor activity (PTFC) by high resolution analytical methods was performed. PTFC had a molecular weight of 6,000 dalton by the size-exclusion HPLC analysis; it showed a marked UV-absorbance spot at 254 nm and a fluorescent spot at 366 nm on the thin-layer plate by thin-layer chromatography which spots coincided at the same place of the plate. The amino acid composition and sequencing analyses showed that PTFC consisted of at least twelve different amino acids, but the amino acid sequence could not be determined. The combined results indicate that PTFC is a compound with a molecular weight of 6,000 dalton, composed of peptide and nucleotide-like material. The peptide is rich in aspartic acid and its N-terminal end may be blocked.

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.