Extraction of [14CI Vitamin a from Rat Liver and Kidney During Processing for Transmission Electron Microscopy

The retention of radio isotope-labekd vitamin A during processing for electron microscopy was investigated using the livers and kidneys of vitamin A deficient rats. [15-¹⁴C]Retinol (3μCi/animal) was administered by esophageal intubation to male rats which had been maintained on a vitamin A deficient diet for five or sir weeks postweaning. Glutaraldehyde- or osmium-fixed tissue was processed by three methods: a) routine (a graded series of ethanols, propylene oxide and epoxy), b) rapid (75% and 95% ethanol with three changes of epoxy), or c) water-soluble embedding (70% and 80% hydrorypropyl methacrylate). Water-soluble embedding retained the highest percentage of label in the tissue (liver: 96.31%; kidney: 98.68%). Inclusion of osmium tetroxide in the processing sequence and minimal exposure of tissue to lipid solvents were necessary for good retention of labeled vitamin A in tissues.

The opinions or assertions contained herein are the private views of the author and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense.

In conducting the research described in this report, the investigators adhered to the “Guide for Laboratory Animal Facilities and Care,” as promulgated by the Committee on the Guide for Laboratory Animal Resources, National Academy of Sciences—National Research Council.     

Extraction of [14C] vitamin A from rat liver and kidney during processing for transmission electron microscopy.

The retention of radioisotope-labeled vitamin A during processing for electron microscopy was investigated using the livers and kidneys of vitamin A deficient rats. [15-14C]Retinol (3muCi/animal) was administered by esophageal intubation to male rats which had been maintained on a vitamin A deficient diet for five or six weeks postweaning. Glutaraldehyde- or osmium-fixed tissue was processed by three methods: a) routine (a graded series of ethanols, propylene oxide and epoxy), b) rapid (75% and 95% ethanol with three changes of epoxy), or c) water-soluble embedding (70% and 80% hydroxypropyl methacrylate). Water-soluble embedding retained the highest percentage of label in the tissue (liver: 96.31%; kidney: 98.68%). Inclusion of osmium tetroxide in the processing sequence and minimal exposure of tissue to lipid solvents were necessary for good retention of labeled vitamin A in tissues.     

The ontogenesis of calcium-binding protein in fetal rat kidney

Summary The presence of calcium-binding protein (CaBP) in the nucleus and cytoplasm of epithelial cells of renal tubules varies with the physiologic state. As part of a study to determine whether or not intranuclear CaBP precedes intracytoplasmic CaBP in the same cell, we used peroxidase-labeled antibody against human renal CaBP to localize CaBP in fetal rat kidney tubules. This paper reports examination of kidneys from rats on each day of gestation from the 10th to term (21 days) and on each of the first seven postnatal days. CaBP was first detected in fetal rat kidneys on the 19th gestational day. The histochemical staining reaction that revealed the CaBP was less intense than that produced in kidneys from adult animals, but its distribution was like that in adults, with some cells having no CaBP, others having it in the cytoplasm only, in the nucleus only, or in both. By the seventh postnatal day the staining reaction was similar to the adult patterns in all respects. Send offprint request to:commander,Department of Comparative Medicine, Letterman Army Institute of Research, Attn. : Research Librarian, Presidio of San Francisco, CA 94129, USAIn conducting the research described in this report, the investigators adhered to the Guide for Laboratory Animal Facilities and Care, as promulgated by the Committee on the Guide for Laboratory Animal Resources, National Academy of Sciences-National Research CouncilThe opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense     

Vitamin A regulates obesity in WNIN/Ob obese rat; independent of stearoyl-CoA desaturase-1

Stearoyl-CoA desaturase 1 (SCD1), an important enzyme involved in monounsaturated fatty acid biosynthesis is a key player in energy homeostasis. Here, we tested the impact of vitamin A on hepatic and adipose tissue SCD1 expression and adiposity per se, using an obese mutant rat strain namely, WNIN/Ob developed at National Center for Laboratory Animal Sciences of National Institute of Nutrition, India. Seven months-old 24 male lean and obese rats of WNIN/Ob strain were divided into two groups; each group was subdivided into two subgroups having 6 lean and 6 obese rats and received diets containing either 2.6 mg or 129 mg vitamin A/kg diet for two months. Feeding of high (but non-toxic) doses of vitamin A resulted in significant reduction in body weight gain, and retroperitoneal white adipose tissue weight (RPWAT) in obese rats. Further, vitamin A feeding resulted in augmented expression of SCD1 in liver and RPWAT of lean rats, while no such effect was seen in obese rats. Taken together, the present data suggest that vitamin A decreases body weight gain in obese rat model independent of SCD1 gene regulation.     

Chronic dietary vitamin A supplementation regulates obesity in an obese mutant WNIN/Ob rat model

Objective: To understand the possible role of chronic dietary high vitamin A supplementation in body weight regulation and obesity using a novel WNIN/Ob obese rat model developed at the National Centre for Laboratory Animal Sciences of National Institute of Nutrition, India. Research Methods and Procedures: Thirty‐six 7‐month‐old male rats of lean, carrier, and obese phenotypes were broadly divided into two groups; each group was subdivided into three subgroups consisting of six lean, six carrier, and six obese rats and received diets containing either 2.6 or 129 mg vitamin A/kg of diet for 2 months. Body weight gain, food intake, and weights of various organs were recorded. Adiposity index and BMI were calculated. Serum and liver retinol and brown adipose tissue (BAT)‐uncoupling protein1 (UCP1) mRNA expression levels were quantified. Results: Chronic feeding of high but non‐toxic doses of vitamin A through diet significantly reduced (P ≤ 0.05) body weight gain, adiposity index, and retroperitoneal white adipose tissue mass (without affecting food intake) in obese rats compared with their lean and carrier counterparts. In general, vitamin A treatment significantly improved hepatic retinol stores (P ≤ 0.05) in all phenotypes without affecting serum free retinol levels. However, augmented BAT‐UCP1 expression was observed only in carrier and obese rats (whose basal expression was low). Discussion: Our data suggest that chronic dietary vitamin A supplementation at high doses effectively regulates obesity in obese phenotype of the WNIN/Ob strain, possibly through up‐regulation of the BAT‐UCP1 gene and associated adipose tissue loss. However, in vitamin A‐supplemented lean and carrier rats, changes in adiposity could not be related to BAT‐UCP1 expression levels.     

Anticonvulsants for soman-induced seizure activity

This report describes studies of anticonvulsants for the organophosphorus (OP) nerve agent soman: a basic research effort to understand how different pharmacological classes of compounds influence the expression of seizure produced by soman in rats, and a drug screening effort to determine whether clinically useful antiepileptics can modulate soman-induced seizures in rats. Electroencephalographic (EEG) recordings were used in these studies. Basic studies were conducted in rats pretreated with HI-6 and challenged with 1.6×LD₅₀ soman. Antimuscarinic compounds were extremely effective in blocking (pretreatment) or terminating soman seizures when given 5 min after seizure onset. However, significantly higher doses were required when treatment was delayed for more than 10 min, and some antimuscarinic compounds lost anticonvulsant efficacy when treatment was delayed for more than 40 min. Diazepam blocked seizure onset, yet seizures could recur after an initial period of anticonvulsant effect at doses 2.5 mg/kg. Diazepam could terminate ongoing seizures when given 5 min after seizure onset, but doses up to 20 mg/kg were ineffective when treatment was delayed for 40 min. The GABA uptake inhibitor, tiagabine, was ineffective in blocking or terminating soman motor convulsions or seizures. The glutamate receptor antagonists, NBQX, GYKI 52466, and memantine, had weak or minimal antiseizure activity, even at doses that virtually eliminated signs of motor convulsions. The antinicotinic, mecamylamine, was ineffective in blocking or stopping seizure activity. Pretreatment with a narrow range of doses of ₂-adrenergic agonist, clonidine, produced variable protection (40–60%) against seizure onset; treatment after seizure onset with clonidine was not effective. Screening studies in rats, using HI-6 pretreatment, showed that benzodiazepines (diazepam, midazolam and lorazepam) were quite effective when given 5 min after seizure onset, but lost their efficacy when given 40 min after onset. The barbiturate, pentobarbital, was modestly effective in terminating seizures when given 5 or 40 min after seizure onset, while other clinically effective antiepileptic drugs, trimethadione and valproic acid, were only slightly effective when given 5 min after onset. In contrast, phenytoin, carbamazepine, ethosuximide, magnesium sulfate, lamotrigine, primidone, felbamate, acetazolamide, and ketamine were ineffective.The animals used in studies performed in, or sponsored by, this Institute were handled in accordance with the principles stated in the Guide for the Care and Use of Laboratory Animals, proposed by the Committee to Revise the Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources, National Research Council, and published by National Academy Press, 1996, and the Animal Welfare Act of 1966, as amended. The opinions or assertions contained herein are the private views of the authors, and are not to be construed as reflecting the views of the Department of the Army or the Department of Defense.     

TASTE ACCEPTANCE IN SQUIRREL MONKEYS (SAIMIRI SCIUREUS)

Six squirrel monkeys were presented with solutions representingthe four primary tastes. The solutions included various concentrationsof glucose or sodium saccharine (sweet), sodium chloride (salty),citric acid (sour), and quinine sulfate or sucrose octaacetate(bitter). A 24 hr two-bottle choice technique was employed.Amount of food, water, and solution consumed every 24 hr wasrecorded. The results showed that the maximum intake for glucosesolution was with the 5.0% concentration, although maximum caloricintake was with the 1.25% concentration where there was a potentiationof food intake. Water was preferred over sodium saccharine atthree of the four concentrations which were tested, and waterwas preferred over or equally to the concentrations of sodiumchloride and citric acid that were used. However, quinine sulfateand sucrose octaacetate were preferred over or equally to waterat most of the concentrations which were tested. In conducting the research described in this report, the investigatorsadhered to the ‘Guide for Laboratory Animal Facilitiesand Care’, as promulgated by the Committee on the guidefor Laboratory Animal Resources, National Academy of Sciences- National Research Council.     

Ultrastructural studies of muscle cells and vascular endothelium immediately after freeze-thaw injury

Little is known about the ultrastructural changes which occur in vascular endothelium immediately after an in vivo freezethaw insult, although most investigators will agree that tissue viability relates directly to the degree of vascular damage. In this study an electron microscopic examination of an in vivo model for frostbite injury was initiated. The horseradish peroxidase technique was utilized to follow early alterations in capillary flow and the independent effects of hypoxia, cooling to 2 °C, supercooling, and a single freeze-thaw insult were assessed. No precipitous changes in muscle cell mitochondria or capillary endothelium were detected as a result of brief hypoxia, cooling at 2 °C, or supercooling to ?13 °C. Reducing the temperature by 1 °C/min until freezing occurred, continuing to cool for 10 min postheat of fusion, and rapidly rewarming resulted in consistent mitochondrial damage in muscle cells and marked degeneration of associated capillaries. Peroxidase injected iv prior to thawing was rarely localized in the capillaries of previously frozen muscle. Since peroxidase was found in the capillaries of unfrozen legs of the same animals, it is inferred that little or no flow occurred in most capillaries postthaw. Ultrastructural integrity of capillaries immediately after thawing may be a good index for predicting tissue loss.“In conducting the research described in this report, the investigators adhered to the ‘Guide for Laboratory Animal Facilities and Care,’ as promulgated by the Committee on the Guide for Laboratory Animal Facilities and Care of the Institute of Laboratory Animal Resources, National Academy of Sciences-National Research Council.”     

Immune reconstitution prevents metastatic recurrence of murine osteosarcoma

Primary tumors developing in immunocompetent hosts escape immunosurveillance by acquiring immune evasive properties. This raises the prospect that metastases derived from such tumors will also evade immunity. To investigate whether immune surveillance plays a role in preventing metastases, we studied a murine model which mimics the clinical progression of osteosarcoma: primary tumor growth in the lower extremity, amputation, minimal residual disease followed by the development of overt metastases. K7M2 implants readily escaped immune surveillance since normal BALB/c mice, T cell deficient SCID and T/NK cell deficient SCID-bg mice showed no difference in the rate of growth of primary osteosarcomas. However, both SCID and SCID-bg mice had higher rates of metastases than immunocompetent mice. Similarly, immune reconstitution following transfer of naive T cells to SCID or SCID-bg mice did not impact primary tumor growth, but significantly diminished metastatic recurrence. T cells in osteosarcoma bearing mice produced IFNγ in response to tumor and IFNγ production by immune reconstituting T cells was required to prevent metastases. These results demonstrate an important role for T cell based immune surveillance in preventing metastases, even when metastases develop from tumors that adeptly evade immunosurveillance. The results further suggest that T cell depleting cancer therapies may eliminate beneficial immune responses and that immune reconstitution of lymphopenic cancer patients could prevent metastatic recurrence of solid tumors. By acceptance of this article, the publisher or recipient acknowledges right of the U.S. Government to retain a nonexclusive, royalty-free license in and to any copyright covering the article. The contents of this publication do not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. Animal care was provided in accordance with procedures outlined in the “Guide for the Care and Use of Laboratory Animals” (NIH Pub. No. 86-23, 1996). This project was funded in whole or part with funds from the National Cancer Institute, National Institutes of Health, under Contract No. NO1-CO-56000.     

A serological comparison of the three morphological phases of coccidioides immitis: Complement fixation tests with a Pooled antiserum obtained from rabbits with experimental Coccidioidomycosis

Summary A sonicated spherule preparation was more reactive than a sonicated arthrospore antigen in complement fixation tests with a pooled serum from rabbits with experimental coccidioidomycosis. The reactivity of the sonicated spherule approximated the reactivity of coccidioidin. When the sonicated spherule was separated into its supernatant and sediment fractions, both preparations exhibited serological activity.

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Zusammenfassung Ein mit Schallwellen hergestelltes Kügelchen-Präparat war activer in dem Komplement-Tests mit Blutserum von Kaninchen mit einer Coccidioidomycosisinfektion als ein mit Schallwellen hergestelltes Arthrosporantigen. Die Reaktivität der mit Schallwellen hergestellten Kügelchen war der des Coccidioidin ähnlich. Wurden diese Kügelchen in Niederschlag und Lösung getrennt, so hatten beide Präparate serologische Aktivität.

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Part of a dissertation submitted to the Graduate School of Duke University in partial fulfillment of requirements for the Ph.D. degree.This work was supported by contract with the Department of the Army, Fort Detrick, Frederick, Maryland.In conducting the research reported herein, the investigators adhered to Guide for Laboratory Animal Facilities and Care established by the Committee on the Guide for Laboratory Animal Facilities and Care of the Institute of Laboratory Animal Resources, NAS—NRC.     

Culture of cells in beem capsules: A new technique for electron microscopic study of monolayer cultures

Summary A method is described for the monolayer cultivation of primary cell suspensions and established cell lines directly in carbon-coated BEEM capsules. BEEM capsules are routinely employed by electron microscopists in tissue embedding procedures; growing monolayer cultures directly on the lids of inverted BEEM capsules presents the obvious advantage of maintaining cell to cell to substratum contacts with a minimum of stress and damage in the preparative steps for electron microscopy. This work was supported by grant AM 17631 from the National Institute of Arthritis, Metabolism, and Digestive Diseases, grant CA 11339 from the National Cancer Institute. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

New Books

Beletsky , L. D. & Orians , G. H. 1996: Red-winged blackbirds: decision-making and reproductive success. Houck , L. D. & Drickamer , L. C. (eds) 1996: Foundations of animal behavior: classic papers with commentaries. Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council 1996: Guide for the care and use of laboratory animals.     

Development of dome epithelium in gut-associated lymphoid tissues: Association of IgA with M cells

Summary The dome epithelium (DE), which covers gutassociated lymphoid tissues (GALT) and provides both a protective barrier over lymphoid follicles and a route for antigen uptake from the gut, develops in rabbit appendix (caecum) during the first week of neonatal life. To determine if secretory immunoglobulins from maternal milk interact with this developing tissue, their interrelationships in neonatal rabbit appendix were examined by use of immunocytochemical techniques. The glycoprotein, secretory component, was not produced by neonatal rabbits less than 15 days old, since neither the membranous nor the free, secreted forms of maternal secretory component were associated with villi or DE of neonates. Immunoglobulin A (IgA), but neither IgG nor IgM, were noted on DE by light microscopy, even though IgG was abundant in the villus lamina propria and vascular spaces. The epithelial IgA was distributed, in a patchy pattern, across the upper dome surface of some two-day-old, and all five-and ten-day old nursing animals, but IgA was not on DE of rabbits prevented from nursing. Immunoelectron microscopy of appendix from nursed rabbits revealed IgA directly over the apical surface of M cells, where it formed a continous, thick coating without binding to adjacent immature absorptive cells; it was also within apical vacuoles of M cell cytoplasm. The distribution of IgA on the DE of rabbit appendices indicated that in differentiating GALT, maternal IgA reacted preferentially with M cells or pre-M cells, leading to speculation concerning a role for IgA in the development of GALT and in establishment of mucosal immune responses in neonates.In conducting the research described in this report, the investigators adhered to the standards set forth in the Guide for the Care and Use of Laboratory Animals (NIH Publication 85-23) as promulgated by the Committee on Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources, National Research Council, USAAbbreviations DE dome epithelium - GALT gutassociated lymphoid tissues - HRP horseradish peroxidase - IgA immunoglobulin A - SC secretory component The views of the authors expressed here do not purport to reflect the position of the Department of the Army or the Department of Defense     

The cultivation of the exoerythrocytic stages ofPlasmodium berghei from sporozoites

Summary Cultures of embryonic rat brain and liver, and embryonic turkey brain were inoculated with sporozoites ofPlasmodium berghei. Sporozoites succeeded in establishing exoerythrocytic infections in approximately 10% of the cultures. The exoerythrocytic parasites developed to a late schizont stage with some showing early segmentation although free merozoites were not observed. The morphology and rate of development of the exoerythrocytic parasites in culture appear similar to that seen in vivo. This work was supported by ONR Contract No. N00014-76-C-1132 and Naval Medical Research and Development Command, Research Work Unit No. M0095PN.002.5058. the opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large. The experiments reported herein were conducted according to the principles set forth in theGuide for the Care and Use of Laboratory Animals, Institute of Laboratory Resources, National Research Council, DHEW, Pub. No. (NIH) 74-23.     

Rhesus monkey micro mixed lymphocyte and mitogen reactivity: Optimal conditions and normal variability

Optimal conditions for the rhesus monkey micro mixed lymphocyte system with multiple automated harvesting of samples were evaluated. Parameters studied were cell concentration, length of culture period, methods of inactivation of cell populations, supplementation of media, type of culture plates, and changes in the reactivity of cells from individual animals over an extended time period. This work was supported in part by Portland Veterans Administration Hospital, Portland, Oregon, and the General Research Support Branch of the U.S. Public Health Service Grant RR00163, the Bureau of Medicine and Surgery, Navy Department, Work Unit No. M4318. 01.007ABG2. The opinions or assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the U.S. Navy Department or the Naval service at large. The animals used in this study were handled in accordance with the provisions of Public Law 89–54 as amended by Public Law 91–579, “Animal Welfare Act of 1970,” and the principles outlined in the “Guide for the Care of Laboratory Animals,” U.S. Department of Health, Education, and Welfare Publication No. (NIH) 73-23.     

Precocious development of lectin (Ulex europaeus agglutinin I) receptors in dome epithelium of gut-associated lymphoid tissues

Summary Dome epithelium (DE), the tissue covering lymphoid domes of gut-associated lymphoid tissues, was examined in both adult and neonatal rabbit appendix or sacculus rotundus to determine if dome epithelial cells matured earlier than epithelial cells covering adjacent villi. The localization of well-differentiated epithelial cells in rabbit gut-associated lymphoid tissues (GALT) was accomplished histochemically by use of molecular probes: fluorescein isothiocyanate or horseradish peroxidase conjugates of Ulex europaeus agglutinin I (UEA), a lectin specific for terminal L-fucose molecules on certain glycoconjugates. The villus epithelial cells of newborn and 2-, 5-, or 10-day-old rabbits did not bind UEA, but between the twelfth and fifteenth days of postnatal life, UEA receptors were expressed by well-differentiated villus epithelial cells. In contrast to villus epithelium, DE in appendix and sacculus rotundus of neonatal rabbits expressed UEA receptors two days after birth, a feature that distinguished the DE of neonatal GALT for the next two weeks. In adult rabbits, UEA receptors were associated with dome epithelial cells extending from the mouths of glandular crypts to the upper domes; in contrast to the domes, UEA receptors were only present on well-differentiated epithelial cells at the villus tips. Results suggested that in neonatal rabbits most dome epithelial cells developed UEA receptors shortly after birth, reflecting precocious development of DE as compared to villus epithelium. In adult rabbit dome epithelium UEA receptors appeared on dome epithelial cells as they left the glandular crypts, representing accelerated epithelial maturation.Abbreviations DE dome epithelium - DEL dome epithelial lymphocytes - FITC fluorescein isothiocyanate - HRP horseradish peroxidase - PBS phosphate-buffered saline - PBS-CaMg PBS containing calcium and magnesium ions - UEA Ulex europaeus agglutinin I The views of the authors expressed here do not purport to reflect the position of the Department of the Army or the Department of DefenseIn conducting the research described in this report, the investigators adhered to standards set forth in the Guide for the Care and Use of Laboratory Animals (NIH Publication 85-23) as promulgated by the Committee on Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources, National Research Council, USA     

Morphological changes in the junctional complex of cells in the arachnoid layer of the rat after cold injury

Summary The junctional complexes of cells in the outer arachnoid layer overlying the cerebral cortex of 2-week-old rats were examined with freeze-fracture electron microscopy up to 60 min after transcranial cold injury to the dorsal surface of the brain. Within 30 min after injury, areas of gap and tight junctions with morphological features characteristic of junction formation and/or junction disruption were found scattered among normal junctional complexes in some arachnoid cells. Within 60 min after injury, tight junctions with features typical of less leaky zonulae occludentes were present in all arachnoid cells examined. These morphological features include increases in the number of tight junctional strands and the number of strand-to-strand anatomoses. Gap junctions were interspersed among the tight junctional strands, and many were completely encircled by the strands. The increase in the number and complexity of the tight junctional strands in response to brain injury may be the morphological basis for the maintenance of the cerebrospinal fluid-blood dural barrier.This study was supported by the National Institute of Neurological and Communicative Disorders and Stroke Grant NS20590. The opinions or assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the DoD or the USUHS. The experiments reported herein were conducted according to the principles set forth in the Guide for Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Research Council, DHEW Pub. No. (NIH) 78-23     

Embedding of Neural Tissue in Agarose or Glyoxyl Agarose for Vibratome Sectioning

Agarose was used to embed the brain or spinal cord of lampreys or rats before cutting vibratome sections. Agarose embedding was compatible with immunocytochemistry or the use of horseradish peroxidase as a neuroanatomical tracer. Concentrated agarose with high intrinsic gel strength was optimal for embedding glutaraldehyde fixed neural tissue. A quick procedure was to blot tissue and embed in 5% (w/v] Sigma type I-A or Litex type LSL agarose at 45-55 C dissolved in 50 mM neutral-pH TFUS buffer before cutting 50-100 μm vibratome sections. An alternative procedure that improved retention of tissue sections in the agarose was to rinse the tissue in H₂0, blot and embed in 5% (w/v] Sigma type I-A or Litex type LSL agarose at 45-55 C dissolved in H₂0, then equilibrate the block overnight in buffer. Phosphate buffer prevented complete dissolving of agarose. Tissue could be covalently linked to the embedding matrix using a novel aldehyde-derived agarose (NuFix® FMC BioProducts). Slices of spinal cord from neonatal rats could be cut after embedding in 5% FMC Seaprep® agarose in rat Ringer's at 23-26 C.     

A serological comparison of the three morphological phases of Coccidioides immitis; complement fixation tests with anti-Histoplasma capsulatum serum

Summary The results of this study indicated that antigens prepared from the three morphological phases ofCoccidioides immitis differed in their complement fixing activity with anti-Histoplasma capsulatum pooled serum. Spherule antigens were serologically less active in tests with the anti-H. capsulatum pooled serum than antigens prepared from arthrospores and from mycelium.Antigenic determinants which are common toC. immitis andH. capsulatum appeared to be located on the intact arthrospore cellular surface but not on the surface of spherule cells.Part of a dissertation submitted to the Graduate School of Duke University in partial fulfillment of requirements for the Ph. D. degree.This work was supported by contract with the Department of the Army, Fort Detrick, Frederick, Maryland.In conducting the research reported herein, the investigators adhered to Guide for Laboratory Animal Facilities and Care established by the Committee on the Guide for Laboratory Animal Facilities and Care of the Institute of Laboratory Animal Resources, NAS-NRC.     

Lack of expression on cultured human T-lymphocytes of a T-cell antigen shared by the molt-4 cell line and normal human thymocytes

Summary Four hematopoietic cell lines (CCRF-CEM, HSB-2, MOLT-4, and RPMI-8402), derived from acute lymphoblastic leukemia and expressing T-cell surface markers (T-HCL), were studied with two specific anti-T-cell sera. The sera were raised in rabbits against human thymocytes (anti-HTY) and against T-cells cultured in the presence of conditioned medium derived from lymphocytes stimulated with PHA (anti-CTC). Both sera were absorbed to obtain a T-cell specific pattern of reaction and were further absorbed with normal peripheral blood lymphocytes or with each of the four T-HCL. The anti-HTY sera absorbed with CEM, 8402, and HSB-2 still reacted with MOLT-4. A similar pattern of reactivity was found only with the anti-CTC absorbed with 8402, whereas, after absorptions with the other cell lines, this antiserum was unreactive against MOLT-4. After absorption with normal peripheral blood lymphocytes, anti-HTY still reacted with thymocytes and MOLT-4 but was negative on CTC. In contrast, anti-CTC absorbed with peripheral blood lymphocytes (PBL) was negative on thymocytes and MOLT-4 but still reacted against CTC. Our data confirm the existence of a T-cell antigen (probably an early T-cell differentiation antigen) shared between thymus and MOLT-4. This antigen is not expressed on CTC, although these cells express an antigenic pattern more complex than PBL. Antisera to CTC represents a source of anti-T-cell sera free of contamination with antibodies to early thymus-related antigens but containing other T-cell-related specificities. Supported in part by Naval Medical Research and Development Command, Research Task No. ZF51.524.013.1025, and National Cancer Institute Contract No. Y01-CB-00319. The opinions and assertions contained herein are the private ones of the writers and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large. The experiments reported herein were conducted according to the principles set forth in the current edition of the “Guide for the Care and Use of Laboratory Animals,” Institute of Laboratory Animal Resources, National Research Council.