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Populations of 90 Uromyces appendiculatus isolates were collected from throughout the Americas and evaluated for virulence on 19 standard bean rust differentials, and also on 12 landraces of Phaseolus vulgaris from South and Central America. The landrace differentials represented geographical centers of bean domestication. Three groups were observed. Two groups were isolates from centers of bean domestication and a third heterogeneous group comprised isolates from countries in South and Central America. Molecular analysis using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) was also conducted on these isolates. Cluster analysis of the molecular profiles showed three groups that corresponded to those obtained by virulence tests. These results show a clear differentiation of the pathogen population along similar lines as its host and suggest parallel evolution in the bean rust pathosystem.

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We have investigated the ontogeny of MHC class I, class II, CD45, and macrophage antigens in whole mounts of normal human fetal retina at 10-25 weeks gestation (WG) using monoclonal antibodies and immunogold histochemistry. MHC class I antigens were expressed on retinal vascular endothelial cells and provided a useful marker of vessel organization from 14-25 WG. Microglial cells expressed immunoreactivity to MHC class I, class II, and CD45 antigens from 10 WG (pre-vascularization) and macrophage S22 (Mac S22) antigen from 14 WG (post-vascularization), although none of the antigens tested were detected on neuronal or macroglial elements. Microglia expressing MHC, CD45, and macrophage antigens occurred in both ramified and rounded forms with no close correlation being observed between morphology and antigenicity. The numbers of immunoreactive cells labeled with each of the four markers increased steadily throughout gestation in all specimens studied. Equivalent numbers of microglia expressed MHC class I, class II, and CD45 antigens in retinae at similar gestational ages; however, our data indicate that microglia expressing Mac S22 antigen comprise approximately 40% or less of the population of MHC and CD45-immunoreactive cells during development. Topographical analyses suggest that MHC class I, class II, and CD45-positive microglia enter the retina from both the peripheral retinal margin and the optic disc from at least 10 WG; Mac S22-positive cells appear in association with the development of the retinal vasculature and enter the retina via the optic disc after 14 WG.

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The development of microglial topography in wholemounts of human retina has been examined in the age range 10-25 weeks gestation (WG) using histochemistry and immunohistochemistry for CD45 and major histocompatibility complex class II antigens. Microglia were present in three planes corresponding to the developing nerve fibre layer/ganglion cell layer, the inner plexiform layer and the outer plexiform layer. Distribution patterns of cells through the retinal thickness and across the retinal surface area varied with gestational age. Microglia were elongated in superficial retina, large and ramified in the middle plane, and small, rounded and less ramified in deep retina. Intensely labeled, rounded profiles seen at the pars caeca of the ciliary processes, the retinal margin and at the optic disc may represent precursors of some retinal microglia. At 10 WG, the highest densities of microglia were present in middle and deep retina in the far periphery and at the retinal margin, with few superficial microglia evident centrally at the optic disc. At 14 WG, high densities of microglia were apparent superficially at the optic disc; microglia of middle and deep retina were distributed at more central locations although continuing to concentrate in the retinal periphery. Microglia appear to migrate into the developing human retina from two mains sources, the retinal margin and the optic disc, most likely originating from the blood vessels of the ciliary body and iris, and the retinal vasculature, respectively. The data suggest that the development of microglial topography occurs in two phases, an early phase occurring prior to vascularization, and a late phase associated with the development of the retinal vasculature.

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PURPOSE: To investigate the distribution and phenotype of major histocompatibility complex (MHC) class II-positive dendritic cells and macrophages in normal human fetal cornea in the age range 10 to 25 weeks gestation. METHODS: Peroxidase and gold immunohistochemistry were used to visualize MHC class II and macrophage antigen (S22) immunoreactive cells. Cell distributions were analyzed quantitatively, and topographic maps were produced. RESULTS: Immunoreactive cells, concentrated centrally, were present at 10 weeks gestation in the corneal epithelium and stroma. Average densities increased steadily up to 25 weeks gestation. Two morphologic forms of MHC class II and S22 immunoreactive cells were observed--large, dendritiform cells and small, rounded cells with short processes. Electron microscopy revealed that most MHC class II-positive cells were morphologically consistent with previous ultrastructural descriptions of corneal Langerhans cells. Immunoreactive cells were more numerous in immunogold-labeled specimens than in peroxidase-labeled specimens of similar ages. However, quantitative analysis of both techniques revealed that S22-positive cells comprised 30% to 50% of MHC class II-positive cells. CONCLUSIONS: This study provides a detailed description of heterogeneous populations of MHC class II and S22 immunoreactive cells in the human fetal cornea. In contrast to the adult cornea, which is typically devoid of MHC class II-positive cells, immunoreactive cells in the fetal cornea are concentrated centrally and increase in density up to at least 25 weeks gestation. These results indicate that reduction in Langerhans cell numbers to adult levels must occur after 25 weeks gestation. The presence of dendritic cells and macrophages in the fetal cornea has important implications for the understanding of corneal immunology.

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NADPH-diaphorase histochemistry was used to identify and analyse the topography of cones in adult human retinae (in the age range 44 to 68 years). Retinae were treated for NADPH-diaphorase reactivity, present in vascular walls, amacrine cells and cone and rod outer segments; a subpopulation of less intensely reactive profiles, morphologically resembling cones, possibly represents the blue cone population. Regularly spaced pairs of cones (one intensely labelled and one weakly labelled cone) were also apparent throughout the retina, and were most common along the horizontal meridian, particularly towards the periphery. The diameters and distributions of labelled cone outer segments were assessed using image analysis. Cone density in the adult retina ranged from 2000 per mm2 in the temporal periphery, to 82,000 to 120,000 per mm2 at the fovea centralis. Distribution patterns confirmed the presence of a cone streak, extending from the foveal region into nasal retina, but no evidence of superior-inferior asymmetry was detected.

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