Blood Cells Of Monkeys Research Articles

ABO typing in experimental cynomolgus monkeys using non-invasive methods

ABH antigens are not expressed on the red blood cells of monkeys, making it difficult to accurately determine their blood type. In this study, we evaluated the feasibility, convenience, and stability of two non-invasive methods for ABO typing (a reverse gel system assay and a buccal mucosal cell immunofluorescent assay) in cynomolgus monkeys (n = 72). The renal tissue immunofluorescent assay was used to obtain an accurate blood type in the monkeys. Using the reverse gel system assay and preabsorbed serum, we achieved accurate detection of ABO blood groups in 65 of the 72 monkeys but obtained confusing results in the remaining 7. The original immunofluorescent staining of the buccal mucosal smears clearly and correctly identified the ABO blood groups in 50 of the 72 monkeys. After repeated smearing and staining, the ABO group type could be correctly identified in samples from the rest of the monkeys, which were either lacking sufficient buccal mucosal cells or contained impurities. Based on our findings, we recommend the reverse gel system assay as the first choice for primate blood type analysis, and the buccal mucosal cell immunofluorescent assay as a Supplementary Method whenever the reverse gel system assay fails to give a clear result.

The In Vivo Effects of Recombinant Human Interleukin-3: Demonstration of Basophil Differentiation Factor, Histamine-Producing Activity, and Priming of GM-CSF-Responsive Progenitors in Nonhuman Primates

Recently human interleukin-3 (IL-3) produced by molecular cloning was characterized as a growth factor for basophils and eosinophils in human bone marrow cultures. Since we found a similar activity of the human factor on simian bone marrow cells, we investigated the in vivo effects of recombinant human (rh) IL-3 in healthy rhesus monkeys (n = 10). rh IL-3 was administered subcutaneously (SC) to monkeys at different doses (11, 33, and 100 micrograms/kg/d) for 14 days followed by subsequent rh GM-CSF administration (5.5 micrograms/kg/d SC) for another two weeks. During the second week of rh IL-3 administration monkeys responded with a twofold to threefold increase of WBCs caused by a dose-dependent elevation of basophils (up to 40% of WBCs) and eosinophils. rh IL-3 also induced a dose-dependent increase of histamine (up to 700-fold above normal values) in monkey blood cells. Administration of rh GM-CSF to rh IL-3 pretreated monkeys resulted in a twofold enhanced increase in WBCs (due mainly to eosinophils and neutrophils) compared with animals treated with rh GM-CSF alone. Simultaneous administration of both cytokines (100 micrograms/kg rh IL-3 + 5.5 micrograms/kg rh GM-CSF SC) to two separate monkeys for 14 days induced a WBC elevation similar to that observed in monkeys treated with rh GM-CSF alone. In conclusion, our results indicate that rh IL-3 is a differentiation factor for blood basophils and primes the hematopoietic system for subsequent rh GM-CSF actions.

The in vivo effects of recombinant human interleukin-3: demonstration of basophil differentiation factor, histamine-producing activity, and priming of GM-CSF-responsive progenitors in nonhuman primates

Recently human interleukin-3 (IL-3) produced by molecular cloning was characterized as a growth factor for basophils and eosinophils in human bone marrow cultures. Since we found a similar activity of the human factor on simian bone marrow cells, we investigated the in vivo effects of recombinant human (rh) IL-3 in healthy rhesus monkeys (n = 10). rh IL-3 was administered subcutaneously (SC) to monkeys at different doses (11, 33, and 100 micrograms/kg/d) for 14 days followed by subsequent rh GM-CSF administration (5.5 micrograms/kg/d SC) for another two weeks. During the second week of rh IL-3 administration monkeys responded with a twofold to threefold increase of WBCs caused by a dose-dependent elevation of basophils (up to 40% of WBCs) and eosinophils. rh IL-3 also induced a dose-dependent increase of histamine (up to 700-fold above normal values) in monkey blood cells. Administration of rh GM-CSF to rh IL-3 pretreated monkeys resulted in a twofold enhanced increase in WBCs (due mainly to eosinophils and neutrophils) compared with animals treated with rh GM-CSF alone. Simultaneous administration of both cytokines (100 micrograms/kg rh IL-3 + 5.5 micrograms/kg rh GM-CSF SC) to two separate monkeys for 14 days induced a WBC elevation similar to that observed in monkeys treated with rh GM-CSF alone. In conclusion, our results indicate that rh IL-3 is a differentiation factor for blood basophils and primes the hematopoietic system for subsequent rh GM-CSF actions.

Chromosomal aberrations and DNA repair ability of in vitro irradiated white blood cells of monkeys previously exposed to total body irradiation

Six monkeys ( Macaca fascicularis) were total-body-irradiated with 60Co (fractionated irradiation of 8 or 10 Gy). Blood samples were collected at different times post total-body irradiation, then in vitro irradiated in order to test whether a prior in vivo irradiation could affect the radiosensitivity of their leukocytes. We suggested in a preliminary report that the enhanced chromosomal radiosensitivity of in vivo irradiated monkeys could be correlated with a DNA repair deficiency (Guedeney et al., 1986). chromosomal aberrations, the rate of initial strand breaks and their rejoining estimated using a fluorescent assay for DNA unwinding were chosen as the endpoints in this more extensive study. We observed that the yield of dicentrics induced by a subsequent in vitro irradiation was lower than that scored in unirradiated monkeys in few cases (6/22) whereas the number of acentrics was found to be modified in 16 of the 22 samples. An altered DNA repair ability was observed in most but not all blood samples tested. Thus, in view of such intra-individual variability, the results of this more extensive study lead us to conclude that a previous total-body irradiation does not alter the γ-induced chromosome aberrations and DNA repair ability in a reproducible manner.

自動血球分類装置MICROXの実験動物への応用

Automatic blood cell analyzer "MICROX" was examined for possible applicability to the differential count of white blood cell in the monkey and in the dog. Spun blood smear slides prepared from 42 monkeys and 60 dogs were stained with Wright's dye and totals of 4971 white blood cells of the monkey and 7189 cells of the dog were counted, respectively. The identification rate of MICROX was checked by the cell by cell counting method on a monitor TV of the instrument. Analysis of the data was made to examine correlationship between the results of MICROX and of optical microscopy on the same slides. 1) Identification rates and misclassification rates were 97.0% and 3.0% respectively in monkeys and 96.4% and 3.6% in dogs. 2) A high degree of correlation was observed between the results by MICROX and by optical microscopy in respect of segmented neutrophils and lymphocytes. 3) There were an average of 15.5 cells counted as unknown cells in monkey blood and 16.5 in dog blood per 100 cells, respectively. Of the unknown cells about 40 to 50% were closely disposed leukocytes within the same counting squares and almost all of atypical lymphocytes and erythroblasts were also counted as unknown cells on the analyzer. The data support the feasibility of the use of MICROX for the differential count of white blood cells of normal monkeys and dogs.

Structure, hemagglutinins, and subgroup position of adenovirus 20, 25, and 28.

By ultracentrifugation and red cell adsorption experiments it was proved, that hemagglutinating activity of adenovirus types 20, 25, and 28 against monkey blood cells is associated with the virus particles; while no soluble hemagglutinins were found for type 20 and 25, the soluble hemagglutinating activity of type 28 against rat blood cells was located in a dodecon. The length of the fibres, as observed on the virus surface or in pentons, was 115 to 120 A for type 20, 25, and 28, coinciding with a reference virus (type 9–15) of subgroup II.

The nature of monkey cell haemagglutinins of Rosen's group II adenoviruses.

Some of the adenovirus type of Rosen’s group II exhibit weak agglutinating properties for monkey blood cells. The relation of these haemagglutinins to haemagglutinins for rat blood cells and to the infectious virus was studied by cross-absorption with blood cells and by density gradient centrifugation with caesium chloride. In the virus types studied (type 9, 13, and a wild strain of type 15) one part of the monkey cell haemagglutinin was associated with the virion and, for type 9 and 15, was inseparable from the virus-associated haemagglutinin for rat blood cells. In density gradient centrifugation a second haemagglutinin for monkey blood cells was found, separable from the virus particles and associated with the portion of higher buoyant density of the soluble haemagglutinin for rat blood cells. In type 13 and 23, haemagglutinins for monkey and rat blood cells can be separated by absorption with the heterologous blood cell species. Haemagglutinins for monkey cells are resistant to trypsin or trypsin plus subsequent heating to 55°C, in contrast to haemagglutinins for rat blood cells. Monkey cell haemagglutination is reversibly inhibited by the presence of CsCl. In haemagglutination-inhibition tests it was found that haemagglutination with monkey blood cells is more difficult to be inhibited by immune serum that haemagglutination with rat blood cells.

Glucose-6-phosphate dehydrogenase activity in the red blood cells of monkeys.

IN human beings there exists an inborn character of the red blood cells which is usually harmless, but which, in certain individuals, can lead to severe hæmolysis if the red cells are challenged by the ingestion of certain drugs or foods. The main abnormality in these individuals is a deficiency of the enzyme glucose-6-phosphate dehydrogenase in the red cells, which usually have a correspondingly low amount of reduced glutathione in them. Further, when such sensitive red cells are incubated in certain chemicals, for example, acetylphenylhydrazine, there is a marked fall in the level of reduced glutathione, which does not occur with erythrocytes of normal persons. This is the glutathione stability test of Beutler1, and it has been pointed out by Beutler et al.2 that the glutathione instability of the drug-sensitive cells probably results from their deficiency in glucose-6-phosphate dehydrogenase. A simple test for demonstrating this enzyme deficiency state has been developed by Motulsky3.

Sodium Salicylate Inhibiting Anti-Rh Immunization in Animals

SummaryIn guinea pigs and in rabbits the formation of anti-Rh agglutinins following the injection of rhesus monkey blood cells appears to be diminished when sodium salicylate has been administered 3 days prior to and during the period of immunization.