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Biomedical subjects

B M Gebhardt

Publications and source records attributed to B M Gebhardt.

87 records · Page 5Linked to original sources
PubMed1974

Clonal character of F1 hybrid lymphocyte subset recognition of parental cells in one-way mixed lymphocyte cultures.

Proliferation of F(1) hybrid lymphocytes in mixed lymphocyte cultures is stimulated by mitomycin-blocked parental cells. The demonstration of this phenomenon using F(1) hybrids derived from congenic lines of mice establishes that the stimulation is controlled by genes in or closely linked to the major histocompatibility locus chromosome region. In agreement with the finding that tumor-bearing mice have an increased capacity for primary alloantigen recognition, it was observed that the F(1) hybrid response to parent was also augmented by tumor bearing. Chromosomal analysis of dividing cells in one-way mixed cultures confirms that F(1) cells, and not the blocked parental cells, enter mitosis. Stimulation of F(1) cells by a soluble mediator liberated by the parental cells was not observed and mitomycin blocking of parental cells seems to be a completely effective blocking agent ensuring that parental cells can not enter DNA synthesis. The specificity and clonal nature of F(1) recognition of parent was demonstrated using a 5-bromodeoxyuridine-suicide procedure. Distinct clones of lymphocytes in F(1) spleen cell populations seem to recognize one or the other parent, but not both, in such experiments. These observations and others in tumor systems suggest that most or all heterozygous organisms may possess potentially self-reactive clones of lymphocytes.

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PubMed1966

Evidence from cultured leucocytes of blood cell chimerism in ex-parabiotic frogs.

Postmetamorphic diploid and triploid frogs that had earlier been joined in parabiotic union from embryonic life until metamorphosis were each found to be chimeric with respect to their blood cells, as revealed in chromosome preparations of cultured leucocytes. Blood cells precursorsmost likely were interchanged when the ex-parabionts shared a common circulation in embryonic life,and the exchanged precursor cells apparently homed in the hematopoietic tissues of the hosts. The tolerance which exparabiotic pairs of frogs exhibit toward grafts of each other's skin is attributable to the blood cell chimerism.

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PubMedDate not supplied

Bioavailability of microsphere-entrapped cyclosporine A in the cornea and aqueous of rabbits.

Extended release of immunosuppressive drugs and sustained drug levels are desirable for the treatment of corneal graft rejection and other ocular immune disorders. This experiment was conducted with biodegradable microspheres containing cyclosporine A to assess their suitability for achieving this goal. Microspheres containing cyclosporine A were prepared using a solvent evaporation process. A mixture of poly(lactic) and poly(glycolic) acid polymers (50:50) and cyclosporine A was dissolved in a mixture of chloroform and acetone. The solution was then emulsified in an aqueous solution of polyvinyl alcohol and stirred for 24 hours to evaporate the organic solvent. The final assayed concentration of cyclosporine A was 15.38 mg/mL. A 0.13 mL aliquot (2.0 mg) of the suspension of the microspheres was injected subconjunctivally in 24 eyes of white New Zealand rabbits. The concentration of cyclosporine A in the aqueous and cornea was measured at 6, 12, 24, 48, 72, and 144 hours after injection (n = 4 for each group). Corneal levels of cyclosporine A ranged from 2392 +/- 70 ng/mL at 6 hours to 1297 +/- 459 ng/mL at 144 hours. The aqueous levels ranged from 110 +/- 0 ng/mL at 6 hours to 62 +/- 11 ng/mL at 144 hours. These data indicate that a microsphere drug delivery system is an effective means of delivering cyclosporine A to the cornea and anterior chamber, and may provide an alternative for the treatment of ocular immune disorders.

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PubMedDate not supplied

Modulation of in vitro immune reactions by platelet activating factor and a platelet activating factor antagonist.

Platelet activating factor (PAF) was found to suppress primary and secondary mixed lymphocyte reactions and BN52021, a naturally occurring PAF antagonist, blocked PAF-mediated suppression and enhanced the mixed lymphocyte reactions. The effect of delayed addition of PAF or BN52021 24 h or later after the initiation of cultures reduced the suppressive and enhancing effects, respectively. The removal of the antagonist BN52021 from mixed lymphocyte cultures up to 72 h after their initiation also was found to eliminate the potentiating effect of this antagonist. The continuous presence of PAF in mixed lymphocyte cultures used to generate cytotoxic lymphocytes suppressed the generation of effector cells while the addition of BN52021 elicited an enhanced level of cell-mediated cytotoxicity in such cultures. BN52021 enhanced the cytotoxic activity in such cultures irrespective of the presence of exogenous interleukin-2, suggesting that the antagonist-mediated enhancement is not due to the enhanced production of interleukin-2 by cells in the mixed cultures. The experiments reported here provide evidence for a role of PAF in modulating the complex interactions that take place in the initiation of cellular immune reactions. Furthermore, the results of these experiments indicate that the immunoregulatory action of PAF can be modulated by an antagonist that exerts its action independently of the production of the lymphocyte growth factor, interleukin-2.

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PubMedDate not supplied

Reactivation of herpes virus after lamellar keratoplasty.

PURPOSE: To determine if lamellar keratoplasty in rabbits latently infected with herpes simplex virus type 1 (HSV-1) would stimulate graft recipients to shed virus and induce viral-specific corneal lesions. METHODS: Rabbits latently infected with HSV-1 received lamellar allografts in one eye from normal uninfected rabbits and the contralateral eyes served as unoperated controls. Normal rabbits received lamellar grafts from rabbits latently infected with HSV-1. For 1 week after surgery, slit-lamp examination and ocular swab sampling were performed daily to assess viral reactivation. RESULTS: The occurrence of positive swab cultures and corneal epithelial lesions after lamellar keratoplasty was significantly higher in operated eyes of latently infected rabbits when compared to the control eyes. Ocular shedding or recurrent lesions were not observed in the normal rabbits receiving corneal grafts from latently infected donors. CONCLUSIONS: These results indicated that lamellar keratoplasty induces HSV-1 shedding and recurrent epithelial lesions in the eyes of rabbits latently infected with HSV-1, which received lamellar grafts, but not in the eyes of normal rabbits given lamellar grafts from HSV-1 latently infected rabbits. It seems that the site of viral latency is not the anterior corneal stroma or the epithelium.

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PubMedDate not supplied

Immunogenicity of epikeratophakia tissue lenses containing living donor keratocytes.

The purpose of this study was to compare the survival of epikeratophakia tissue lenses prepared with cryolathing and lyophilization (frozen lenses) and without (fresh lenses) in donor-sensitized recipients with vascularized corneas. Fresh lenses placed in vascularized corneas of immune recipients were subjected to immune attack. Frozen lenses placed in vascularized corneas of immunized recipients did not elicit an immune reaction. Neither fresh nor frozen lenses elicited immune reactions in nonvascularized corneas of immune recipients or in nonvascularized, nonimmune recipients. These results indicate that although the fresh lenses are more antigenic than the lenses in which the cells have been killed by freezing and lyophilization, the fresh lenses prepared using the BKS-1000 technique containing living stromal keratocytes are not likely to stimulate allograft immune reactions in unsensitized patients with avascular graft sites.

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