Baboon Blood Research Articles

The ligand for c-kit, stem cell factor, stimulates the circulation of cells that engraft lethally irradiated baboons

Recombinant human stem cell factor (SCF), the ligand for c-kit, has been shown to stimulate increased numbers of hematopoietic progenitor cells of multiple types to circulate in the blood of baboons, but it was not known if the cells stimulated to circulate by SCF contained cells capable of engrafting and rescuing lethally irradiated baboons. Peripheral blood mononuclear cells (PBMNC) were collected by leukapheresis from four untreated control baboons and from three baboons on the 10th or 11th day of treatment with SCF (200 micrograms/kg/d). All animals were transplanted with 1.00 to 1.04 x 10(8)/kg of cryopreserved autologous PBMNC after treatment with a single dose of 1,020 cGy total body irradiation (TBI). Three animals were transplanted with PBMNC that had been collected during SCF treatment, 24 to 38 days after the last dose of SCF. Rapid trilineage engraftment was documented by bone marrow biopsy in all three. The mean time to a total white blood cell count (WBC) > or = 500/microL, WBC > or = 1,000/microL, and an absolute neutrophil count (ANC) > or = 500/microL was 15 +/- 3 (mean +/- SD), 19 +/- 1, and 19 +/- 2 days, respectively. Two animals remain alive with stable engraftment more than 180 and 245 days posttransplant. The third died of sepsis 32 days posttransplant with a hypercellular marrow showing trilineage engraftment. The surviving animals were transfusion independent by 10 and 59 days posttransplant. Four control animals were transplanted with PBMNC collected in the absence of SCF stimulation. One was treated for 11 days with SCF (200 micrograms/kg/d) after PBMNC were collected. This animal was transplanted 25 days after the last dose of SCF. None of the four control animals engrafted and they died 13, 16, 28, and 38 days posttransplant with marrow aplasia. Treatment with SCF stimulates the circulation of cells that engraft and rescue lethally irradiated baboons. The characteristics of the transplantable cells present in the circulation are now amenable to direct study.

The Ligand for c-kit, Stem Cell Factor, Stimulates the Circulation of Cells That Engraft Lethally Irradiated Baboons

AbstractRecombinant human stem cell factor (SCF), the ligand for c-kit, has been shown to stimulate increased numbers of hematopoietic progenitor cells of multiple types to circulate in the blood of baboons, but it was not known if the cells stimulated to circulate by SCF contained cells capable of engrafting and rescuing lethally irradiated baboons. Peripheral blood mononuclear cells (PBMNC) were collected by leukapheresis from four untreated control baboons and from three baboons on the 10th or 11th day of treatment with SCF (200 µg/kg/d). All animals were transplanted with 1.00 to 1.04 × 108/kg of cryopreserved autologous PBMNC after treatment with a single dose of 1,020 cGy total body irradiation (TBI). Three animals were transplanted with PBMNC that had been collected during SCF treatment, 24 to 38 days after the last dose of SCF. Rapid trilineage engraftment was documented by bone marrow biopsy in all three. The mean time to a total white blood cell count (WBC) ≥ 500/μL, WBC ≥ 1,000/μL, and an absolute neutrophil count (ANC) ≥ 500/μL was 15 ± 3 (mean ± SD), 19 ± 1, and 19 ± 2 days, respectively. Two animals remain alive with stable engraftment more than 180 and 245 days posttransplant. The third died of sepsis 32 days posttransplant with a hypercellular marrow showing trilineage engraftment. The surviving animals were transfusion independent by 10 and 59 days posttransplant. Four control animals were transplanted with PBMNC collected in the absence of SCF stimulation. One was treated for 11 days with SCF (200 μg/kg/d) after PBMNC were collected. This animal was transplanted 25 days after the last dose of SCF. None of the four control animals engrafted and they died 13, 16, 28, and 38 days posttransplant with marrow aplasia. Treatment with SCF stimulates the circulation of cells that engraft and rescue lethally irradiated baboons. The characteristics of the transplantable cells present in the circulation are now amenable to direct study.

A c-kit ligand, recombinant human stem cell factor, mediates reversible expansion of multiple CD34+ colony-forming cell types in blood and marrow of baboons

The ligand for the human c-kit, recombinant human stem cell factor (SCF), was administered to baboons at doses of 200, 100, 50, 25, and 10 micrograms/kg/d. SCF induced a dose-dependent expansion of hematopoietic colony-forming cells (CFC) of multiple types in both blood and marrow, including colony-forming unit (CFU) granulocyte- monocyte, burst-forming unit-erythroid, CFU-MIX, and high proliferative potential-CFC. These changes were associated with a dose-dependent leukocytosis, involving all leukocyte lineages, a reticulocytosis, and increases in marrow cellularity. At 200 micrograms/kg/d of SCF, CFC in blood were increased 10-fold to greater than 100-fold. This correlated with an increased frequency of CD34+ cells in blood. The frequency of CFC in blood approached that of marrow in some animals. These changes were reversed within 7 to 14 days of stopping SCF. The results of these studies suggest a role for the c-kit ligand in stimulating the expansion of multiple CFC types in blood and marrow for potential therapeutic purposes.

A c-kit Ligand, Recombinant Human Stem Cell Factor, Mediates Reversible Expansion of Multiple CD34+ Colony-Forming Cell Types in Blood and Marrow of Baboons

The ligand for the human c-kit, recombinant human stem cell factor (SCF), was administered to baboons at doses of 200, 100, 50, 25, and 10 micrograms/kg/d. SCF induced a dose-dependent expansion of hematopoietic colony-forming cells (CFC) of multiple types in both blood and marrow, including colony-forming unit (CFU) granulocyte-monocyte, burst-forming unit-erythroid, CFU-MIX, and high proliferative potential-CFC. These changes were associated with a dose-dependent leukocytosis, involving all leukocyte lineages, a reticulocytosis, and increases in marrow cellularity. At 200 micrograms/kg/d of SCF, CFC in blood were increased 10-fold to greater than 100-fold. This correlated with an increased frequency of CD34+ cells in blood. The frequency of CFC in blood approached that of marrow in some animals. These changes were reversed within 7 to 14 days of stopping SCF. The results of these studies suggest a role for the c-kit ligand in stimulating the expansion of multiple CFC types in blood and marrow for potential therapeutic purposes.

Plasma pharmacokinetics and metabolism of the benzodiazepine antagonist [11C] Ro 15-1788 (flumazenil) in baboon and human during positron emission tomography studies.

Flumazenil is a specific antagonist of the central benzodiazepine receptor (CBZR). Labelled with 11C, this compound is the reference radioligand for positron emission tomography (PET) study of the CBZR in humans and primates. The time-course of [11C]-flumazenil radioactivity and its main acid metabolite [11C] Ro 15-3890 were reconstructed from the time-course of total radioactivity in plasma after administration with high or low SRA in primates and humans, applying an extraction procedure validated by TLC. The measured pharmacokinetics of [11C]-flumazenil (T1/2 beta = 45.1 +/- 12.3 min, T1/2 alpha = 1.5 +/- 1.5 min; K = 0.14 +/- 0.14 min-1; Vd area = 44.0 +/- 17.0 l; Clp = 40.0 +/- 8.5 l/h) exhibited a very rapid distribution phase followed by fast elimination, with a large volume of distribution; these results were confirmed by HPLC determinations and agree with previously published data on unlabelled flumazenil. Pharmacokinetics of [11C] Ro 15-3890 acid metabolite show that high drug concentrations in the blood are promptly achieved (kf = 0.13 +/- 0.004 min-1), with a very rapid elimination half-life (T1/2m = 4.47 +/- 1.31 min) comparable to that of [11C]-flumazenil. The percentage metabolization of parent compound to the acid [11C] Ro 15-3890 was constant from the 15th minute and was significantly higher in man compared to the monkey. This percentage was increased by prior eating. The other putative metabolites, i.e. labelled [11C] Ro 15-4965 and unlabelled Ro 15-5528, were never observed at detectable concentrations with TLC and HPLC in rabbit, baboon and human blood samples. This pharmacokinetic study of plasma flumazenil may be useful to implement a dynamic method of CBZR quantification using PET and for analysis of pharmacokinetics in brain tissue.

A comparison of methods for the separation of [ 11C]Ro 15-1788 (flumazenil) from its metabolites in the blood of rabbits, baboons and humans

The determination of the percentage of unchanged [ 11C]Ro 15-1788 with respect to total radioactivity, essential for the pharmacokinetic exploitation of data of blood radioactivity decrease obtained in PET studies, may be carried out with equal results using either an extraction procedure or a TLC method. The analysis of blood samples of various sources (rabbits, baboons, healthy volunteers and anxious patients) and the use of an automatic linear scanner to quantify the radioactive distribution on the TLC plates lead to the conclusion that only [ 11C]Ro 15-1788 and its acid metabolite [ 11C]Ro 15-3890 and no other radioactive compound are present in blood at detectable concentration.

A baboon model for pregnancy-associated antigens (PAPP-A, PP5, PP14)

By radioimmunoassays established on human derived antigens, PAPP-A, PP5 and PP14 immunoreactivity was detected in placental extracts and blood of pregnant baboons. None of the serial dilution curves suggested parallelism between respective human and baboon samples. Based on slopes of regressed logit-log transformed binding data, PAPP-A demonstrated the greatest degree of interspecies immunological crossreactivity. PP14 showed the least conservation of antigenic determinants. Physicochemical characterization on heparin, zinc chelate and bovine thrombin affinity matrices could not distinguish human from baboon-derived antigens. As in the human, baboon PAPP-A and PP5 were not detected in blood of male or non-pregnant animals. PP14 was detected in baboon follicular fluid, and only PP5 immunoreactivity was measured in culture media of baboon embryos. Of the three antigens, PAPP-A was detected in pregnant baboons at about 61 days gestation, that is, 4 weeks before PP5 and PP14. With the exception of PP14 which attained peak concentration at 118 days of pregnancy, PAPP-A and PP5 concentrations were greatest at term. In conjunction with physicochemical and immunological criteria, these physiological kinetics clearly support a role for developing a baboon model to serve for further studies into feto-maternal signals, particularly antigens such as PAPP-A and PP5.

Formula omitted]: The radioligand of choice for pett studies of the dopamine receptor in human brain

N-methylspiroperidol, the amide N-methyl analogue of the neuroleptic spiroperidol, was radiolabeled with fluorine-18, and its distribution in the baboon brain was studied using positron emission transaxial tomography. Stereospecific binding was demonstrated in the striatum (but not in the cerebellum) by pretreatment with (-)- or (+)-butaclamol. The kinetic distribution was similar to that of [ 18F]spiroperidol, but the absolute striatal uptake (in percent of administered dose) was at least two-fold higher. Analysis of baboon blood at 10 min after injection indicated that less than half of the radioactivity in the plasma was due to unchanged radioligand. Analysis of the metabolic stability of [ 18 F]-N- methylspiroperidol in rat brain for 4 hr indicated that, like [ 18F]spiroperidol, it is very stable to metabolic transformation in the rat central nervous system. Striatal uptake and retention in the rat was five-fold higher for [ 18 F]-N- methylspiroperidol than for [ 18F]spiroperidol. These results suggest that [ 18 F]-N- methylspiroperidol is an ideal choice for studies of the dopamine receptor in humans.

The effects of ritodrine on prostaglandin metabolite concentrations in the blood of pregnant baboons.

To investigate the relationship between beta sympathomimetic drugs and prostaglandins, we measured prostaglandin metabolites in the plasma of pregnant baboons that were given the drug ritodrine. Animals were at a mean gestation of 120 days, which is equivalent to 27 weeks in women. Ritodrine was infused i.v. at a rate of 23 micrograms/min or 80 micrograms/min for 4 h. Plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM), 11-deoxy-13,14-dihydro-15-keto-11,16-cyclo-prostaglandin E2 (PGEM-11), and 6-keto-prostaglandin-F1 alpha (6-keto-PGF1 alpha) were determined as indicators of the precursors, prostaglandin F2 alpha, prostaglandin E2, and prostacyclin (PGI2), respectively. At the 23 micrograms/min ritodrine infusion rate plasma levels of 6-keto-PGF1 alpha were decreased by 51% (+/- 15%) in the animals receiving the drug (p less than 0.005) whereas corresponding levels in control animals did not differ significantly from pre-infusion levels. At the 80 micrograms/min ritodrine infusion rate, the plasma levels of 6-keto-PGF1 alpha decreased by 66% (+/- 18%) compared with pre-infusion levels (p less than 0.001) but again the levels in control animals were not changed significantly. Plasma levels of PGFM and PGEM-11 in animals during ritodrine treatment did not differ significantly from baseline values.

Vitamin fluctuations in the blood of female baboons in relation to normal menstrual cycles, treatments with Lo-Ovral or Depo-Provera and a selected vitamin supplement

Both endogenous and exogenous sex steriods may induce changes in plasma vitamin levels by altering availability, transport, binding, or use of vitamins. This study investigated some of those mechanisms by observing in a primate model (baboon), the blood levels of carotene, folate, vitamins A, B12, and C, and the status of vitamin B6, riboflavin, and thiamin. The latter three vitamins were studied by determining their relationship to asparate aminotransferase, glutathione reductase, and thiamin transketolase, respectively. The vitamin screen was obtained throughout normal menstrual cycles in 10 baboons and weekly for 55 wk in five baboons receiving Lo-Ovral and in four baboons receiving Provera. During the last 16 wk of hormonal treatment, all baboons received a vitamin supplement containing pyridoxine, riboflavin, and thiamin. Only carotene (p less than 0.0001), vitamin A (p less than 0.05), glutathione reductase (p less than 0.05), and thiamin transketolase (p less than 0.05) levels fluctuated significantly during normal menstrual cycles. Long-term treatment with Lo-Ovral and Provera resulted in numerous changes but there were very few differences between the two hormone treatments. Compared to control levels, vitamin C was elevated during treatment while all three enzyme activities were lowered. Vitamin supplementation raised asparate aminotransferase and glutathione reductase activity and the levels of folic acid, vitamin A, and carotene. This study demonstrates that interactions between hormones and vitamins and among vitamins themselves, are complex but it is likely that the treatments used here caused no physiologically significant vitamin alterations.

Radioimmunoassay of somatomedin-C in the baboon (Papio cynocephalus): A comparison of multiple techniques of measurement.

This study was undertaken to assess the nature of somatomedin-C (SM-C) in baboon (Papio cynocephalus) blood and to compare various methods for estimating SM-C concentrations. Parallel dose-response curves were obtained with normal baboon serum, normal human serum, and purified SM-C. Recovery of purified SM-C added to baboon serum over a wide dosage range (n = 17) was 111 ± 12%, with slightly better recovery at higher potencies. Chromatography of normal baboon serum on Sephadex G-200 at neutral pH produced a profile similar to that observed in the human, as did samples chromatographed on Sephadex G-50 in acid. Although the SM-C content in acid chromatographed plasma was approximately 2.5 times higher than in native unprocessed plasma, there was excellent correlation between the values (r = 0.9143, p < 0.0001). The SM-C in baboon plasma which had been preincubated with glycine HCl was approximately twice that of unprocessed plasma, but the correlation between the two methods was excellent (r = 0.9593, p < 0.0001). The correlation between values obtained after simple acid-ethanol extraction and those observed in unextracted plasma were also significant (r = 0.7689, p < 0.0001). Following a series of four injections of human growth hormone (hGH) to a normal baboon, plasma SM-C rose approximately sevenfold above the initial concentration and returned to basal levels five days after the final injection. These studies show that although the radioimmunoassay (RIA) for SM-C in unprocessed baboon plasma does not measure all of the SM-C present, it provides a reliable index of the total SM-C concentration and reflects GH status in the baboon.

Measuring percent oxygen saturation of hemoglobin, percent carboxyhemoglobin and methemoglobin, and concentrations of total hemoglobin and oxygen in blood of man, dog, and baboon.

We used an automated four-wavelength spectrometer to measure the concentration of total hemoglobin, percent oxyhemoglobin, carboxyhemoglobin, and methemoglobin, and concentration of oxygen bound to hemoglobin in the blood of humans, dogs, and baboons under clinical and various experimental conditions. Measurements of total hemoglobin and methemoglobin with this simple method were comparable to those with standard spectrometric procedures. Carboxyhemoglobin measurements were comparable to those made with gas chromatography, and measurements of oxygen content were comparable to those made with the galvanic cell method. The new instrument is as accurate as the comparison methods used to evaluate it in all parameters, is reliable, and measurements take only 63 s per sample. In addition, it requires minimal operator training, infrequent need for calibration, and no sample preparation.

In vitro studies of cryopreserved baboon granulocytes.

Granulocytes were isolated from the buffy coat of baboon blood by counterflow centrifugation. They were frozen in polypropylene tubes in 2.0 ml volumes containing 1 × 107 granulocytes. The medium consisted of 5% DMSO, 6% HES, 4% human serum albumin, and 6 mM glucose in Normosol‐R, pH 7.1. Granulocytes were cooled to 4 C for 30 minutes, then cooled at 4 C per minute to –80 C and stored for one to three weeks in liquid nitrogen at –197 C. Cooling rates of 1 C and 10 C per minute were less efficacious. Tubes were thawed manually with swirling for 130 seconds at 42 C in a water bath. The yield after thawing was 98 ± 14 per cent.Granulocytes isolated from fresh blood all fluoresced green when incubated with fluorescein diacetate (FDA). Less than one per cent of cells showed nuclear uptake of ethidium bromide (EB). After freezing and thawing, an average of 79 ± 8 per cent granulocytes fluoresced green with FDA and 21 ± 8 per cent fluoresced red with EB. About 98 ± 2 per cent of the granulocytes isolated from fresh blood exhibited a serum dependent capacity for ingestion of latex and yeast. Thawed granulocytes were diluted 1:4 dropwise with a medium containing 7% HES, 4% albumin, and 6 mM glucose in Normosol‐R, incubated with fluorescent latex or fluorescent zymosan at 37 C for 30 minutes and then washed twice. After this, 75 ± 5 per cent demonstrated serum‐dependent latex ingestion and 67 ± 8 per cent ingested zymosan.

The febrile responses in rabbits and rats to leucocyte pyrogens of different species.

1. We have investigated the effects on body temperature of rats and rabbits of leucocyte pyrogen derived from the blood of rat, rabbit, ox, pig and baboon. 2. In the rabbit intravenous injections (3.5 ml.) of solutions containing leucocyte pyrogen derived from ox, pig and rabbit blood produced fevers with short latencies; no fevers resulted from injections of similar solutions derived from rat or baboon blood. 3. In the rat intraperitoneal injections (2.0 ml.) of solutions containing leucocyte pyrogen derived from ox blood caused a fever, while pig leucocyte pyrogen produced a marked hypothermia. Neither rabbit, baboon, nor rat leucocyte pyrogen had any significant effect on rectal temperature of the rats. 4. Our results show that there is a variability of response in the rat and the rabbit to injection of leucocyte pyrogen of different species; leucocyte pyrogen may be species specific.

Stimulation of fetal hemoglobin synthesis in baboons by hemolysis and hypoxia.

Fetal hemoglobin (Hb F) levels in the peripheral blood of baboons (Papio cynocephalus) increased from an average value of 0.78% to 18.1% during the recovery phase from phenylhydrazine-induced hemolytic anemia. A similar increase was observed in animals exposed to hypobaric hypoxia. Large individual variations in the maximal Hb F levels were observed which could not be correlated with the ages of the animals. Reinduction of hemolysis in two fully recovered animals resulted in Hb F levels that were of similar magnitude as in the preceding episode, suggesting the possibility of genetically determined individual variations in the rate of Hb F synthesis under the same conditions of erythropoietic stimulation. Reticulocytes from the animals subjected to hemolysis of hypobaric hypoxia synthesized similar absolute quantities of Hb F in vitro. The results of the present studies indicate that the physiological switch from the synthesis of Hb F to that of Hb A during ontogeny can be reversed in adult nonhuman primates by conditions of erythropoietic stress known to be associated with high erythropoietin levels. These findings open the possibility that Hb F synthesis in adult humans may be therapeutically modulated in individuals who might benefit from increased levels of Hb F, such as patients with sickle cell anemia.

Blood groups of Baboons. Population genetics of feral animals.

AbstractBlood and saliva from 495 Ethiopan baboons were collected in the field and tested for their human‐type A‐B‐O groups while 493 blood samples were tested for their simian‐type blood factors Ap, Bp, Cp, Gp, Np, ca and hu. Four series of feral animals were tested: 194 olive baboons, a troop of 82 and another of 90 hamadryas baboons, and a series of 129 baboons classified as olive/hamadryas hybrids. In addition, 126 baboons from other sources were tested for their human‐type A‐B‐O groups and 131 for their simian‐type blood groups.Human‐type groups A, B and AB but not group O were found in combined series of 621 animals. Gene frequency analysis also indicated the absence of group O. Population analysis of the data obtained for the 493 Ethiopian baboons has shown that the simian‐type blood groups Ap and Bp are independent of one another. In contrast Bp and Gp appear to be determined by corresponding allelic genes; if confirmed by population data on additional series of animals, this would define the first baboon blood group system found. There is a close association between the blood group specificities ca and hu, the exact nature of which still remains to be clarified. Blood group ca, originally believed to be species specific, is found to be polymorphic in both olive baboons and hamadryas as well as in the hybrids; hu, on the other hand, present in all hamadryas tested, is polymorphic only in olive baboons.

Demonstration of Hemopoietic Stem Cells in the Peripheral Blood of Baboons by Cross Circulation

Abstract Baboons were given 1200 R total body irradiation from two opposing 60Co sources. Three animals were given supportive therapy only and died, as expected, within 8 days of irradiation with profound marrow hypoplasia. Five baboons were cross-circulated with unirradiated partners and died within 14 days with evidence suggestive of graft-versus-host disease. Their marrows were repopulated with hemopoietic precursor cells, and three of the five had rises in peripheral white blood cell counts to more than 1500/cu mm before death. These results are compatible with the presence of hemopoietic stem cells in the peripheral blood of a nonhuman primate, the baboon.

Demonstration of hemopoietic stem cells in the peripheral blood of baboons by cross circulation

Baboons were given 1200 R total body irradiation from two opposing 60Co sources. Three animals were given supportive therapy only and died, as expected, within 8 days of irradiation with profound marrow hypoplasia. Five baboons were cross-circulated with unirradiated partners and died within 14 days with evidence suggestive of graft-versus-host disease. Their marrows were repopulated with hemopoietic precursor cells, and three of the five had rises in peripheral white blood cell counts to more than 1500/cu mm before death. These results are compatible with the presence of hemopoietic stem cells in the peripheral blood of a nonhuman primate, the baboon.

Specific resistance of blood at body temperature II.

The specific resistance values for samples of goat, sheep, camel, monkey, baboon, cat and pig blood at 37°C are given for a range of packed-cell volumes extending from 0 to 70%. The relationship is presented in the form p=AeαH, when p is the specific resistance, A and α are constants and H is the packed-cell volume in percent. In addition, values for the change in resistivity with the addition of saline are presented to allow electrical calibration for conductivity cells when saline is used as the indicator in cardiac-output studies.

Early Functional Hemodynamic Impairment in Baboons after 1000 R or Less of Gamma Radiation as Revealed by Hemorrhagic Stress

The present study demonstrated that the existence of radiation-induced impairment of cardiovascular function is disguised by a large hemodynamic reserve in nonhuman primates. These findings were revealed when the cardiovascular function of gamma-irradiated baboons was challenged with three transient bleeding episodes. Thirty baboons in five groups were treated as follows: one group was sham irradiated and the remaining four exposed to either 250, 500, 750, or 1000 R of single, whole-body gamma radiation (35 R/min). The cardiovascular system of each group was challenged at 2, 48, and 168 hr postirradiation with a bleeding episode which lasted approximately 10 min. Each challenge consisted of the rapid withdrawal and replacement of an estimated 43% (25 ml blood/kg body weight) of the baboon's blood volume. Hemodynamic function was assessed before and 30 min after each bleeding episode. The hemodynamic response to the first challenge appeared to be independent of radiation exposure. The greater variability i...