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.

Dialyzable lymphoid extract (DLE) from mice resistant to STZ-induced diabetogenesis can interrupt the progress of diabetes in STZ-treated CD-1 mice.

DLE was prepared from the minority of euglycemic CD-1 mice, previously injected with STZ, and was administered to hyperglycemic CD-1 male mice 1, 2 and 3 weeks after completion of multidose STZ. Mice treated with DLE derived from 2 x 10(7) (IX) or 10(8) lymphocyte equivalents (lymph.equ) were significantly less hyperglycemic than the saline treated controls (P < 0.001). The effects of DLE remained evident for more than 10 weeks after the final DLE treatment. Mice treated with DLE prepared from diabetic mice (hg DLE) developed a somewhat more rapid onset of hyperglycemia than the STZ treated control animals, although this effect did not achieve statistical significance (P = 0.1). This DLE was absorbed on a rat insulinoma cell line (RIN), which contains interspecies cross-reacting islet antigens, and compared to the unabsorbed DLE. Mice treated with hg DLE preabsorbed on RIN cells, showed a slower onset of hyperglycemia. DLE prepared from euglycemia mice and the RIN-absorbed fraction were equally capable of preventing hyperglycemia (P < 0.05). In order to determine whether the DLE effects were genetically restricted, DLE was prepared from BALB/c mice, normally resistant to the diabetogenic effects of multidose STZ, both before and after STZ treatment. STZ primed CD-1 mice treated with 3 weekly doses of 2 x 10(7) lymph. equ. of untreated BALB/c derived DLE, STZ treated BALB/c derived DLE, and STZ treated CD-1 DLE were all less hyperglycemic than the control mice, who received saline (P < 0.001). However, mice treated with CD-1 DLE were less hyperglycemic than the mice given BALB/c derived DLE (P < 0.05). These effects were relatively long-lived. Mice that were given the > 3,500 Dalton fraction of CD-1 DLE were significantly less hyperglycemic than either the control mice or those treated with the < 3,500 Dalton fraction of CD-1 DLE (P < 0.05). Effects remained evident for more than 3 months after the last dose of DLE. Pancreatic tissue from the mice treated with the > 3,500 Dalton fraction of CD-1 derived DLE revealed slightly more islets of a slightly greater size with less surrounding inflammation than either control mice or mice treated with the < 3,500 Dalton fraction of DLE.

Fatal nonpowder firearm wounds: case report and review of the literature.

A previously unreported fatality due to a head wound from a CO2-powered BB pistol is described. In a review of the English language medical literature (primarily forensic), ten previous reports of fatality due to nonpowder firearms were found. A common mechanism is identified in the head wounds leading to fatality. Modern day technology has elevated the BB gun from toy to weapon. Often appearing trivial, BB and pellet gun injuries must be considered in the same class as those from small-caliber low-velocity powder firearms. A patient with a nonpowder firearm injury must be evaluated with a high index of suspicion for injuries that are not apparent during a general physical examination.

A controlled trial of bovine dialyzable leukocyte extract for cryptosporidiosis in patients with AIDS.

Cryptosporidial infection causes severe diarrheal disease in patients with AIDS. Fourteen patients with AIDS and symptomatic cryptosporidiosis were treated with a specific bovine dialyzable leukocyte extract (immune DLE) prepared from lymph node lymphocytes of calves immunized with cryptosporidia or a nonspecific (nonimmune) DLE prepared from nonimmunized calves. Six of 7 patients given immune DLE gained weight and had a decrease in bowel movement frequency, with eradication of oocysts from stool in 5 patients. Six of 7 patients given nonimmune DLE showed no decrease in bowel movement and 4, no clearing of oocytes from stool; 5 continued to lose weight. Subsequently, 5 of these 7 were treated with immune DLE; 4 had a decrease in bowel movement frequency and significant weight gain, with eradication of oocytes from stool in 2 patients. Immune DLE produces sustained symptomatic improvement in patients with AIDS and active cryptosporidiosis, but lack of an appropriate cryptosporidial antigen allows only postulation that an augmentation of cellular immunity to Cryptosporidium parvum induced by immune DLE resulted in the microbiologic and clinical improvement observed.

Treatment of cryptosporidiosis with oral bovine transfer factor.

Cryptosporidia are intestinal protozoans long known to cause diarrhea in humans, especially those with acquired immune deficiency syndrome (AIDS). When transfer factor prepared from calves which possessed delayed-type hypersensitivity to Eimeria bovis was given to nonimmune calves and mice it conferred protection against clinical infection (coccidiosis). Recent studies with oral bovine transfer factor have shown that it can confer cell-mediated immunity to humans. Based on these findings we decided to treat eight AIDS patients suffering from Cryptosporidium-associated diarrhea with transfer factor prepared from calves immune to Cryptosporidium. Prior to treatment with transfer factor, three patients had been treated with spiramycin, one patient with alpha-difluoromethylornithine (DFMO), and one patient with furazolidone for greater than 1 month without clinical or laboratory improvement. Following administration of transfer factor, five or eight patients exhibited a decrease in the number of bowel movements and the development of formed stools. Cryptosporidium was eradicated from the stools of four patients but two of these patients subsequently relapsed and one patient continued to have diarrhea despite the absence of Cryptosporidium in the stool. One patient has been free of diarrhea and Cryptosporidium for 2 years after discontinuation of transfer factor therapy.

Emergency department management of the intoxicated adolescent.

It is widely recognized that many young people will have experiences with alcohol and with psychoactive recreational drugs before emerging from the adolescent years. It is estimated that before leaving ninth grade, one half of the total student population will have had experience with alcohol, and approximately one third with marijuana. Cocaine also is more available to adolescents. For these reasons, the emergency physician is more likely to see teenage patients for drug-related problems.

Juvenile laryngeal papillomatosis with pulmonary spread. Regression following transfer factor therapy.

A 6-year-old girl with a history of juvenile laryngeal papillomatosis since 6 months of age and progressing pulmonary extension of the tumor for two years was treated with transfer factor prepared from her mother. Within one month of the onset of therapy, she exhibited marked clinical improvement. A computed tomographic scan performed after four months of therapy revealed almost complete resolution of her pulmonary lesions.

Deletion of antigen-specific activity from leukocyte dialysates containing transfer factor by antigen-coated polystyrene.

We have reported finding antigen-specific activity in human leukocyte dialysates (DLE) containing TF in the leukocyte migration inhibition (LMI) assay. To analyze this activity further, we have used polystyrene bound to antibody or to antigen as immunoadsorbent for DLE before pulsing nonimmune cells in the LMI assay. Candida-(CAN) immune or diphtheria toxoid-(TOX) immune DLE were depleted of all antigen-specific activity after absorption with specific antigen but not affected by absorption with specific antibody, respectively, and depletion of activity with antigen was abrogated by coating bound antigen with specific antibody before absorption of DLE. CAN-immune, TOX-immune DLE was selectively depleted for either CAN activity or TOX activity after absorption with CAN- or TOX-coated polystryrene, respectively, retaining its CAN-activity when absorbed with TOX and conversely retaining its TOX activity when absorbed with CAN; thus the antigen-specific activity binds to related but not unrelated antigen. The polystyrene-bound antigen-specific activity could be recovered by treatment with 8 M urea. We interpret these findings to suggest that such antigen-specific activity may be either a dialysable fragment of a T cell antigen receptor site, or a portion of the V-region, or a unique Ir gene product that assists in antigen presentation to other T cells.

A method for eliciting indurated DTH reactions to soluble protein antigens in the flank skin of mice: correlation of visual measurements with 125I-UdR uptake indices.

A new technique for eliciting specific indurated delayed type hypersensitivity (DTH) reactions to soluble protein antigens in the flank skin of mice is reported here. We show that immunization with soluble antigens emulsified in complete Freund's adjuvant emulsion allows elicitation of indurated reactions upon intracutaneous test of sensitized mice with antigen on alum precipitates. With this method it is possible to measure simultaneously the gross induration of the cutaneous DTH reactions and correlate it with the degree of mononuclear cell infiltration as quantitated by the cPM of 125I-UdR uptake in mononuclear cells at biopsied reaction sites and confirmed by histological examination.

Antigen-specific activity of murine leukocyte dialysates containing transfer factor on human leukocytes in the leukocyte migration inhibition (LMI) assay.

We report on the extension of the direct leukocyte migration inhibition (LMI) test as an assay for antigen-specific activity in human leukocyte dialysates (DLE) containing transfer factor to an evaluation of antigen-specific activity in DLE prepared from inbred mice. Murine DLE was observed to cause antigen-dependent and antigen-specific effects on the inhibition of migration of nonimmune human leukocyte populations. Pulsing of nonimmune human leukocyte with DLE preparations from BALB/c and SJL mice immunized with Candida, diphtheria toxoid, and SK-SD resulted in their inhibition of migration in the presence of the respective antigens. The antigen-specific activity in murine DLE was found to be present in lymph node cell preparations and to be absent from spleen cell preparations of the same donors. The activity of DLE in lymph node cells was found to be present in the theta-cell enriched subpopulation of nonadherent lymphocytes after passage through nylon wool columns. The antigen-specific activity of murine DLE, as we have reported for human DLE, was found to reside in the < 3500 dalton dialysis fraction and not in the < 3500 dalton fraction. We conclude that nonimmune human leukocytes in the LMI test provide a suitable assay for the detection of antigen-specific activity in murine DLE as well as that in human DLE. Additionally, murine DLE is active across species barriers and appears to share properties with human DLE.

Interaction of subpopulations of murine lymph node lymphocytes in antigen-induced [14C]-thymidine incorporation: T and B cell synergy in the response to antigen.

The antigen-induced [14C]-thymidine incorporation of murine lymph node cells (LNC) that were non-adherent (NAD) or adherent (AD) to nylon wool was studied. In contrast to NAD-LNC, AD-LNC responded like unfractionated LNC, and these responses were T lymphocyte dependent. By co-culturing NAD-LNC with subpopulations of AD-LNC the cellular requirements and interactions necessary for maximal incorporation of [14C]-thymidine were determined. A synergistic effect was observed when NAD-LNC and AD-LNC were co-cultured. Synergism was not dependent on T lymphocytes or macrophages in the AD-LNC population but was associated with the B lymphocyte subpopulation. These results indicate that the number of B lymphocytes present in a population of LNC can significantly influence the magnitude of the response to antigen.

Development of antigen-induced proliferative responsiveness by murine lymph node cells. I. Identification of differences in the in vitro proliferative responses during a first and a second period of responsiveness.

The development and course of antigen-induced proliferative responsiveness by murine lymph node cells (LNC) was observed for 16 weeks post-immunization. The initial phase of responsiveness was characterized by antigen-induced proliferative responsiveness in vitro which reached a maximum 3-5 weeks post-immunization and then declined to low levels by 6-8 weeks. Without injection of additional antigen, the initial phase of responsiveness was followed by the development of a second phase of antigen-induced proliferative responsiveness 10-12 weeks post-immunization. These findings suggest that the in vivo development of lymph node lymphocytes capable of a proliferative response to antigen is under some type of modulation which is maximal 6-8 weeks post-immunization. Early in the first phase the proliferative responses to higher concentrations of antigen peaked early in the culture period (days 3-4), whereas responses to the lower concentrations of antigen were optimal after 5-6 days of culture. During the latter half of the first phase, however, peak proliferative responses were made to all the concentrations of antigen on the same day of culture (day 6). In contrast, the responses detected at the beginning and throughout the second phase of responsiveness were characterized by maximum proliferation to all the concentrations of antigen late in the culture period (day 7). These results delineate the temporal requirements for maturation of antigen-induced proliferative responsiveness of murine LNC post-immunization and indicate the time interval when optimal responses may be detected.