Non-random migration of CD4 and CD8 T cells: changes in the CD4: CD8 ratio and interleukin 2 responsiveness of efferent lymph cells following in vivo antigen challenge

In this investigation we have examined some of the cellular and molecular changes in efferent lymph that drains from an antigen-stimulated peripheral lymph node. Resting efferent lymph is characterized by a higher percentage of CD4+ cells and consequently, a higher CD4/CD8 ratio than peripheral blood. Following antigen stimulation of a cannulated peripheral lymph node in antigen-primed sheep, the percentage of CD4+ cells in efferent lymph increases above the resting level during days 1, 2 and 3 post antigen stimulation. This is followed on days, 3, 4, and 5 after antigen stimulation by an increase in the percentage of CD8+ cells above the resting level which occurs as the percentage of CD4+ cells returns to the resting level. These changes cause the CD4/CD8 ratio to first increase above the resting value during the CD4 phase and then decrease below the resting value during the CD8 phase. During the CD4 phase a lymphokine activity is present in cell-free lymph fluid. Lymph fluid collected at this time supports the proliferation of activated T cells. Supernatants generated from efferent cells collected at a similar time and cultured in vivo for 24 h without any further stimulation are capable of releasing this material. During the CD8 phase cells expressing functional interleukin(IL)2 receptors appear in lymph fluid. The data suggests a sequential exit of T cell subsets from an antigen-stimulated lymph node and that the appearance of IL2-like activity and IL2-responsive cells in efferent lymph fluid are temporally distinct events.     

Studies of efferent lymph cells from nodes stimulated with oxazolone.

Efferent lymph from nodes regional to areas of skin that had been treated with solutions of oxazolone in acetone was collected from unanaesthetized sheep. The application of 5% solutions of oxazolone to unsensitized sheep caused no signs of acute inflammation or ''shut-down'' of lymphocyte traffic; none the less, normal immune responses ensued so that immunoblasts, some containing immunoglobulin, were discharged into the lymph together with specific humoral antibodies. When previously sensitized sheep were challenged with 2.5% solutions of oxazolone the vigorous secondary responses were heralded by Arthus reactions, induced presumably by pre-existing antibodies, which were mainly of the IgG class. A similar sequence of events occurred in a thymus-deprived sheep which had undergone intra-uterine thymectomy at 60 days of gestation. Repeated applications of oxazolone to normal sheep did not exhaust or inhibit the characteristic changes in the flow and composition of the lymph. When immunoblasts from efferent lymph were radiolabeled with 125I-UdR and returned intravenously to the sheep they showed no significant tendency to localize either specifically or non-specifically in areas of skin that had been treated with contact-sensitizing chemicals.     

An outbreak of amebiasis spread by colonic irrigation at a chiropractic clinic

From June 1978 through December 1980, at least 36 cases of amebiasis occurred in persons who had had colonic-irrigation therapy at a chiropractic clinic in western Colorado. Of 10 persons who required colectomy, six did. Of 176 persons who had been to the clinic in the last four months of 1980, 80 had received other forms of treatment. Twenty-one per cent of the colonic-irrigation group had bloody diarrhea, as compared with 1 per cent of the non-irrigation group (P = 0.00013). Thirty-seven per cent of the colonic-irrigation group who submitted specimens had evidence of amebic infection on either stool examination or serum titer, as compared with 2.4 per cent in the non-irrigation group (P = 0.00012). Persons who were given colonic irrigation immediately after a person with bloody diarrhea received it were at the highest risk for the development of amebiasis. Tests of the colonic-irrigation machine after routine cleaning showed heavy contamination with fecal coliform bacteria. The severity of disease in this outbreak may have been related to the route of inoculation.     

Development of a larval bioassay for susceptibility of cat fleas (Siphonaptera: Pulicidae) to imidacloprid

Strategies for controlling cat fleas, Ctenocephalidesfelisfelis (Bouché), have undergone dramatic changes in the past 5 yr. With the advent of on-animal treatments with residual activity the potential for the development of insecticide resistance increases. A larval bioassay was developed to determine the baseline susceptibility of field-collected strains of cat fleas to imidacloprid. All four laboratory strains tested showed a similar level of susceptibility to imidacloprid. Advantages of this bioassay are that smaller numbers of fleas are required because flea eggs are collected for the test. Insect growth regulators and other novel insecticides can also be evaluated. Using a discriminating dose, the detection of reduced susceptibility in field strains can be determined with as few as 40 eggs.     

Peak Identification in Visual Evoked Potentials

Waveform patterns evoked by 4 intensities of flash in normal subjects were studied in relation to intersubject variability. Time-frequency distribution curves of all peaks occurring between 11 and 280 msec after flash onset and meeting minimal criteria were obtained from 46 males. These distributions closely corresponded to similar data reported by others for single intensity stimulation. An algorithm was developed which identified in 67 to 100% of instances a single “peak event’ within the time ranges of each of 6 peak distributions. Many peak events appeared and disappeared within the 4 intensity sets of individuals. Latencies were obtained for these peak events. Application of the algorithm to a replicate sample of 29 Ss, which included 8 females, indicated generalizability. Test-retest data on 15 Ss showed its reliability. The data suggest that methodology significantly contributes to the variability of peak identification among subjects. This may be reduced by employing multiple intensities of stimulation.     

Penetration of teniposide (VM-26) into human intracerebral tumors

Thirty-four consenting patients received VM-26 50–100 mg/m² I.V. before surgical resection of intracerebral tumor, and drug was measured using a high pressure liquid chromatographic technique. Sufficient tumor for analysis was obtained from 29 patients. Brain metastases (13 patients) had higher concentrations of V M-26 than did gliomas (13 patients). Concentrations were comparable in brain metastases and meningiomas (3 patients). Prolonged (24 h) infusion of V M-26 did not appear to result in higher tumor drug concentrations in 5 patients than did rapid (1 h) infusion in 24 patients. Pretreatment with Amphotericin-B 10 mg/m² 12 h and 1 h before VM-26 did not appear to have any effect on VM-26 uptake into 4 intracerebral tumors, although data were limited, and VM-26 concentrations were very high in 1 metastasis. Pretreatment with oral glycerol 500 mg/kg 18 h, 12 h, 6 h, and immediately before I.V. VM-26 may have resulted in increased penetration of VM-26 into 9 tumors, although confirmation is required. Amphotericin-B, glycerol, and operative conditions did not appear to alter VM-26 plasma pharmacokinetics.VM-26 4-demethylepipodophyllotoxin 9-(4-6-O-thenylidene-B-D-glucopyranoside) - VP-16 4-demethylepipodophyllotoxin 9-(4-6-O-ethylidene-B-D-glucopyranoside) Presented in Part at the 74th Annual Meeting of the American Association for Cancer Research, San Diego, California, May 25–28, 183(1).