Biomedical subjects
Publications and source records attributed to D S Gordon.
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A sequential mouse cell culture system is described for the induction and assay of T-helper cells. Unprimed, cortisone-resistant, nylon wool-purified thymocytes cultured with adherent peritoneal exudate cells can be primed in vitro with soluble carrier protein to generate carrier-reactive helper cells. These cultured cells enhance the anti-hapten plaque-forming response of hapten-primed spleen cell cultures to hapten carrier conjugates. The culture conditions, cellular manipulations, and antigen requirements for the optimal induction of helper cells with these purified cell populations is presented. The active helper cell generated in this culture system is a thymus-derived cell which requires macrophages for its induction and must be proliferate in vitro before the manifestation of helper-cell function. Helper cells generated in vitro stimulate both carrier-specific and nonspecific enhancement of splenic anti-hapten responses. The carrier-specific and nonspecific enhancement can be distinguished by the requirement for antigen in the helper cell and spleen cell cultures, the dose of helper cells added to the spleen cell cultures, and by the requirement for additional splenic adherent accessory cell interactions.
Supernates derived from in vitro generated T-helper cells have been analyzed for their capacity to substitute for T-cell carrier reactivity. T-helper cell supernates stimulate both a carrier-specific and nonspecific anti-DNP-PFC response to DNP-carrier conjugates in cultures of hapten-primed spleen cells. The carrier-specific and nonspecific activity can be distinguished by dosage optimum, antigen requirements, binding specificity for carrier, and in the requirement for additional splenic adherent accessory cell involvement. The active factors produced in this system are heat labile and sensitive to trypsin and periodate. They are removed by absorption with alloantisera directed toward the strain from which the supernate was derived but not by a variety of anti-immunoglobulin sera.
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Mononuclear cells (MC) from human blood were fractionated by a variety of physical and immunologic techniques, and the cellular subpopulations generated were assessed for their capacity to lyse herpes simplex virus (HSV)-infected target cells in the presence of IgG antibody to HSV. Latex phagocytosis and surface marker studies were performed in parallel in order to identify the major effector cells by their phagocytic properties and their possession of surface immunoglobulin and receptors for either sheep erythrocytes, C3, or the Fc fragment of IgG. Cytotoxic effector cell activity was unaffected or slightly enhanced after the removal of plastic-adherent or carbonyl iron-adherent MC, indicating that the major effector cell is not a classical monocyte. Similar results were obtained after removal of more than 90% of the T cells by depletion of rosette-forming cells. Likewise, effector cell activity was generally unchanged when more than 95% of the B cells were removed by filtering MC on nylon wool columns. Effector cell function was also found to be normal in three patients with B cell-deficient X-linked agammaglobulinemia. These observations strongly suggest that the effector cells are not T cells or B cells. A 4- to 5-fold enrichment in effector cells, however, was consistently found in a subpopulation, consisting of 5% of the unfractionated MC, that was dramatically enriched both for nonphagocytic cells with only Fc receptor (K cells) and for nonphagocytic cells with no detectable surface markers (null cells). Since, as is demonstrated in the accompanying report, effector surface Fc receptors play a critical role in the mediation of antibody-dependent cellular cytotoxicity directed at HSV-infected target cells, the major mononuclear effector cell in human blood is a K cell.
Thymocyte-macrophage cultures primed with carrier protein release an alloantigen-related factor that enhances the anti-hapten plaque-forming cell response of hapten-primed spleen cells in vitro. Use of immunoadsorbant columns made with a variety of alloantisera indicates that the relevant antigens in this system are coded for the the H-2 major histocompatibility locus in mice. The data indicate that in H-2d and H-2k strains the important genetic regions are in the I region between the K region and the I-C subregion and suggest, based on current understanding of Ia specificities, that the I-A subregion codes for these antigens.
Fluorescent and histochemical methods were applied to the study of the morpho-functional condition of the adrenergic and cholinergic nervous system of the bronchopumonary apparatus in guinea pigs under normal conditions and during sensitization (subcutaneous and inhalant) with tick Dermatophagoides pteronyssinus ticks. There was revealed a marked excitation of the adrenergic nervous system, and a reduction of the acetylcholine esterase activity in the bronchopulmonary tissue during the animal sensitization.
Titanium strips have been used to repair skull defects when preformed metal plates are not avilable and simple wiring would not be enough. The technique is simple, obviates the need to dissect the dura mater, and gives greater resistance to further impact injury than an inlay technique.
For the past few years, the whole blood culture method has been scrutinized for its usefulness and reproducibility in evaluating the immunologic status of patients. This method, used in our laboratory mainly screening specimens for evidence of immunodeficiencies, has been systematically evaluated for reproducibility, and factors affecting normal response through testing specimens from over 70 normal persons. Three mitogens (phytohemagglutinin, Concanavalin A, and pokeweed) were studied; peak time responses occurred later than in the separated cell culture method, and they were different for each mitogen. The dose response curves also depended on the mitogen tested, with pokeweed giving a sharp curve and the others a broad plateau. The harvesting procedure was studied by varying reagents and was optimized for speed and efficiency. The mononuclear cell count of the specimen had little effect on the results as long as it remained less than 3.5 X 10(6)/ml. The results for all mitogens followed a similar distribution curve irrespective of whether the results were expressed as cpm or cpm divided by the mononuclear cell count. The use of the stimulation ratio method to express results was less satisfactory. The mean coefficient of variation for all triplicate samples remained 20% or less and for all conditions tested. In evaluating the immunodeficient patient, the whole blood culture method was found to be equally as informative and easier to perform than the separated cell method.
In summary: (i) the fact that VNS spares auditory function makes it preferable to labyrinthectomy. A comparison of VNS and the "conservative" operations on the inner ear shown that they have much the same effect on hearing. VNS by controlling vertigo may also provide an additional benefit by eliminating one of the stress factors concerned in the perpetuation of hydrops; (2) in the treatment of vertigo, VNS is now the most effective method of treatment. Because of elimination of vertigo is what chiefly concerns most of our patients, we suggest that when medical treatment fails to control vertigo, VNS is the surgical method of choice.
Fluorescein-conjugated staphylococcal protein A (SPA) was complexed with either: 1) heat-aggregated IgG, 2) B cell specific antibody, or 3) T cell specific antibody and then used for an immunofluorescent analysis of mononuclear cell surfaces. Cellular Fc receptors failed to recognize the Fc region of aggregated IgG that had been blocked by SPA. Moreover, fluoresceinated SPA that had been complexed either with anti-Fab (B-cell specific) or T cell-specific antisera prevented the nonspecific binding of these reagents to the IgG-Fc receptors on mononuclear cells, thereby permitting the latter to be properly identified as B or T lymphocytes. In addition, when unconjugated SPA was added to presensitized target cells in a test for antibody-dependent cell-mediated cytotoxicity, cytolysis was abrogated.
The patient with a cerebral gunshot wound has a very unstable condition. In Belfast emphasis has been laid on rapid evacuation and on starting resuscitation within a few minutes of injury. Early and adequate transfusion combats shock; controlled ventilation helps limit the rise in intracranial pressure. Intracranial haematomata should be sought by early operation. Operation seldom improves neurological function in missile wounds of the spine.
Following short-term suspension culture, cells from the Balb/C sarcoma Meth A were allowed to incorporate both [14C] leucine and 2-deoxy-D-glucose-1-[3H] (2DG). The 2DG is trapped as a small anionic marker of the cytosol. Deviation from the kinetics of spontaneous efflux of the markers is interpreted as reflecting perturbation of the target cell membrane. In the presence of guinea pig complement and a rabbit antiserum to Meth A, enhanced 2DG efflux was effected in a titer comparable to that detected with a 51Cr-release assay. With a number of alloantisera and syngeneic immune sera, 2DG efflux was enhanced while 51Cr-release was unaffected. Only in the presence of syngeneic immune sera from mice bearing a low tumor mass, syngeneic splenic leukocytes effect a retardation in the spontaneous 2DG efflux. Sera from animals with a large tumor mass were ineffective. Effux of proteins labeled with [14C] leucine was not altered. The phenomenon was not dependent on the presence of a heat-inactivatable syngeneic complement source. The method described provides a sensitive probe of target cell membrane permeability in the tumor model studied. The phenomenon detected is the capacity of serum, sampled relatively early in syngeneic oncogenesis, to direct syngeneic splenic leukocytes to interact with the target cell membrane differentially altering its permeability to the small cytosol marker.
A patient with dysgraphia resulting from a gunshot wound of the brain is presented. Analysis of the functional status of component operations involved in the skill of writing is described. A retraining scheme based on this analysis brought about a significant improvement in the patient's handwriting.
Falck and Hillarp's method (modified by E.M. Krokhina) was applied to the study of intervertebral ganglia in 45 cats; Nissl's, Unna's methods, and staining for lipofluscin, as well as Masson-Fontana's method for detection of serotonin were used in parallel. Two types of neurons were revealed: a) neurons of the first type with a yellowish-white granularity in the region of the axon hill, dispersed yellowish granules and diffuse greenish fluorescence in the perikarion; b) neurons with an orange granularity accumulated in clustered along the whole territory of the perikarion. Both types of the neurons were surrounded by single adrenergic nerve fibers.
The technique of repairing defects of the skull with titanium is described. The skull contour can be accurately reproduced. The technique is simpler than wiring or suturing methods. The material is inert, radiolucent, and rigid.
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