Mercurochrome Research Articles

Man, monkeys and malaria.

Bizarre though it may now seem, in the last century a whole series of experiments was conducted that involved injecting fresh monkey blood into human volunteers or patients. The reasons, valid at the time, were either to treat neurosyphilis with a relatively benign simian malaria infection (so-called pyrogen therapy), or to establish which monkey malaria species were potential zoonotic reservoirs of infection that then may have interfered with malaria eradication campaigns. Although direct inoculation of fresh blood is the most effective way of retroviruses as well as malaria parasites crossing the species barrier, this hypothesis was never taken up or researched. Unlikely, but not disproved, it is important to remember some of the more hazardous experiments that were done in good faith, too long ago to be recorded on electronic databases.

Characterization of expanded T cell clones in healthy macaques: ontogeny, distribution and stability

Peripheral expanded T cell clones have been discussed mainly in relation to certain diseases or immune function in humans and mice. There is little information on their ontogeny, stability and distribution among T cell subsets as well as major lymphoid organs. We applied reverse transcription-polymerase chain reaction (RT-PCR) with family specific primers for monkey T cell receptor β chain V regions and single-strand conformation polymorphism (SSCP) analysis to analyze the expanded T cell clones in cynomolgus monkeys ( Macaca fascicularis). A number of expanded T cell clones were detected in the peripheral blood of young and adult monkeys, but few expanded T cell clones were detected in the blood of a fetus and a 2-day-old neonate. The clones in adults were maintained over 3 months. These expanded T cell clones were distributed only in peripheral blood and spleen, but few were found in lymph nodes (axillary, inguinal and intestinal). The number of expanded T cell clones was much greater in CD8 single-positive (CD8sp) T cells than in CD4sp T cells, showing that most of these clones originated in the CD8sp T cell population. Almost all the expanded CD8sp T cell clones belonged to the CD28 −, CD29 hi and Fas + subset. The usage of V β genes was not skewed in the 24 V β. Furthermore, higher mRNA signals for effector molecules perforin and IFN-γ were detected in CD8sp T cell subsets with phenotypes of CD28 −, CD29 hi and Fas +, suggesting that the expanded T cells might have developed in relation to T cell activation in the periphery of cynomolgus monkeys.

Quantification of thymic function by measuring T cell receptor excision circles within peripheral blood and lymphoid tissues in monkeys.

The thymus is the primary organ responsible for the production of mature TCR alpha / beta T cells. Quantification of a DNA excision circle that is produced during TCR rearrangement, termed a signal joint TCR rearrangement excision circle (sjTREC) can be used as a measure of thymic function. Here sjTREC measurement has been applied to two monkey species used as animal models of human disease, rhesus macaques (Asian origin) and sooty mangabeys (African origin). Initial PCR analysis determined that the TCR deltaRec-PsiJalpha rearrangement leading to sjTREC formation occurs in both species. Primers to a DNA sequence conserved in macaques, mangabeys and humans were used in a quantitative competitive PCR assay to quantify sjTREC. We found that as in humans, sjTREC in these two monkey species decline with age. sjTREC are first generated in thymocytes during the early stages of TCR rearrangement. Lymph node CD4(+) and CD8(+) T cells contain more sjTREC than peripheral blood T cell populations, suggesting that recent thymic emigrants home to the lymphoid tissues. The sjTREC level is significantly higher within the peripheral blood CD4(+) and CD8(+) T cells of mangabeys compared to macaques. Removal of the thymus in four macaques led to a profound decrease in peripheral blood sjTREC level by 1 year post-thymectomy, indicating the lack of a significant extra-thymic source of peripheral naive T cells in macaques. Our results indicate that production, trafficking, and proliferation of recent thymic emigrants in these two monkey species represents a useful animal model system for understanding human immunological disorders.

Construction of an SIV/HIV type 1 chimeric virus with the human interleukin 6 gene and its production of interleukin 6 in monkey and human cells.

The switch from a Th1- to a Th2-type cytokine response is reported to be involved in human immunodeficiency virus (HIV) disease progression. To study the effect of IL-6, one of the Th2-type cytokines, on AIDS pathogenesis, we constructed an SIV/HIV-1 chimeric virus (SHIV) having the human IL-6 gene (SHIV-IL6) SHIV-IL6 could replicate in M8166, a human T cell line, as well as in monkey and human peripheral blood mononuclear cells (PBMCs). Along with the SHIV-IL6 replication, IL-6 was detected in the culture supernatant by ELISA. The maximum level of IL-6 was 35, 15, and 8 ng/ml in M8166, human PBMCs, and monkey PBMCs, respectively. The expressed IL-6 was biologically active as shown by the proliferation of IL-6-dependent murine hybridoma (MH-60) cells. The inserted IL-6 gene was stable for at least four passages (45 days after the initial infection) in M8166 cells, suggesting the ability to achieve stable expression of IL-6 in long-term experiments. Therefore, we successfully established an SHIV system expressing IL-6, and this is the first report of an SHIV expressing a Th2-type cytokine. With this system, IL-6 should be expressed in the regions where the virus replicates, and therefore the inoculation of macaque monkeys with SHIV-IL6 is expected to provide further information on the etiology of AIDS.

Experimental autoimmune encephalomyelitis in cynomolgus monkeys

Experimental autoimmune encephalomyelitis was induced in macaques. T cell clones infiltrated into the brain lesion area were compared with those in blood. Intradermal immunization of macaques with brain white matter derived from healthy macaque in combination with pertussis toxin, induced neurological symptoms in two macaques. One died on day 25 after immunization, whereas the other survived. Gross examination of the brain from the dead macaque, showed clear hemorrhagic lesions in the white matter. Hematological analysis showed that drastic T cell response was induced in macaques immunized with white matter, but not in control macaques. Flow cytometric analysis of blood cells from the affected macaques demonstrated an increase of CD4 and CD8 T cell populations expressing the CD69 early activation marker. Single strand conformation polymorphism (SSCP) analysis of T cell receptor beta chain showed T cell clones infiltrated into the brain lesion, which were different from those found in the peripheral blood of the same monkey. The present paper shows that SSCP analysis of TCR is useful in studying clonality of T cells infiltrating into the brain tissue of macaque with EAE.

Monkeys of the rainforest in French Guiana are natural reservoirs for P. brasilianum/P. malariae malaria.

Monkey blood samples were collected from 214 monkeys relocated as part of the wildlife rescue organized in French Guiana during the filling of the Petit Saut Dam on the Sinnamary River. These samples were tested for malaria parasites by microscopy of thick blood filsm and by nested PCR for small subunit rRNA genes (SSUrRNA). Parasitic blood forms similar to Plasmodium brasilianum were detected in 4 monkey species: Alouatta seniculus macconnelli, Saguinus midas midas, Pithecia pithecia and Ateles paniscus paniscus, with the highest prevalence in Alouatta monkeys. PCR was more sensitive than the conventional method for detecting low-grade parasitaemia in positive monkeys. The examination of blood films indicated that 5.6% of the animals carried parasites whereas the nested PCR for ribosomal DNA indicated a prevalence of 11.3%. The P. brasilianum SSUrRNA gene sequence was analysed and aligned with those from P. malariae, P. falciparum and P. vivax. This suggested that P. brasilianum and P. malariae are very closely related. Similar results were obtained from analysis of the sequences in P. malariae and P. brasilianum isolates of a polymorphic gene fragment analogous to the merozoite surface protein-1 (MSP-1) gene of P. falciparum. The P. brasilianum/P. malariae sequences were more similar to those of P. vivax than to those of P. falciparum, at least in the gene region examined. The high degree of DNA homology in the sequences of the SSUrRNA and msp1-like genes is consistent with other characterizations demonstrating a taxonomic relationship between P. brasilianum and P. malariae species. Our results provide further evidence that P. brasilianum and P. malariae are virtually identical and should probably be considered to be a single malaria species. This raises the question as to whether monkeys living in the rainforest are natural reservoirs for both simian and human malaria. These results have implications for the interpretation of the current epidemiological situation in French Guiana.

Extrathymic CD4/CD8 double positive T cells

Mature T lymphocytes expressing the αβ T cell receptor are generally classified as either CD4 + or CD8 +, based on the mutually exclusive expression of these two lymphocyte coreceptors. Contrary to this conventional division, there is considerable evidence that significant numbers of CD4/CD8 double positive (DP) lymphocytes exist in the peripheral blood and secondary lymphoid tissues of swine, chickens and monkeys. Although CD4/CD8 DP T cells are rarely present in human peripheral blood the relative percentage of this lymphocyte population can increase spontaneously in healthy individuals and in persons suffering from certain disease conditions. DP can also be found among those T cells infiltrating arthritic joints, rejected kidney grafts and certain tumors. In humans, and rats, CD4/CD8 DP T cells appear transiently following activation of their progenitors. Murine DP cells have been described as a subset of intraepithelial lymphocytes (IELs). However, the relationship of IELs to DP cells in the peripheral blood of other species is unknown. Because of their unconventional phenotype and rarity in human and mice, most immunologists have ignored extrathymic CD4/CD8 DP lymphocytes. Nevertheless, their abundance in the peripheral blood of swine, monkeys and chickens makes it impossible to dismiss this lymphocyte population. Here are reports that have described extrathymic lymphocytes exhibiting a CD4 +CD8 dim phenotype in several species reviewed. Swine and monkey lymphocytes with this phenotype are represented by small resting cells that simultaneously express CD4 and CD8α molecules. The available evidence favors the notion that such DP T cells in swine are comprised predominantly of MHC class II restricted memory CD4 + helper T cells that after activation have acquired the ability to express the CD8α chain and then to maintain this DP phenotype. Moreover, porcine CD4/CD8 DP T cells appear to be comprised of memory cells due to their ability to respond to recall antigen, resilience to thymectomy, increase in proportion with age, expression of memory T cell markers, production of interferon-γ and localization to inflammatory sites. Some of these characteristics are also descriptive of human and monkey CD4/CD8 DP T cells. Thus, in swine, humans and monkeys, these phenotypically distinct lymphocytes appear to represent a primed T cell subset. The possible functional significance of the simultaneous expression of the CD4 and CD8 co-receptors on mature T cells is discussed.

Effect of Genistein on Steroid Hormone Production in the Pregnant Rhesus Monkey

Abstract.Genistein is a phytoestrogen found in soy beans. Phytoestrogens have been reported to cause reproductive problems in sheep and rats. This research was conducted to determine the effects of genistein fed to rhesus monkeys during pregnancy, with specific interest on fetal growth and steroidogenesis in the maternal‐fetoplacental unit. Two groups of five monkeys each were selected in early stages of pregnancy. One group was administered genistein in a fruit treat each weekday until Cesarean section 10 days prior to term. The second, control group, received fruit treats without genistein. Maternal blood samples were collected on Tuesday and Friday of each week. At delivery, samples were collected from the maternal peripheral circulation, uterine veins, uterine‐ovarian veins, and the fetal heart. Comparisons between control and genistein‐treated monkeys revealed no differences in the maternal weight gained during pregnancy, or in fetal weights or placental weights at delivery. Serum was assayed by radioimmunoassay (RIA) for estradiol, progesterone, dehydroepiandrosterone sulfate (DHEA‐S), and estrone. No significant differences (P > 0.05) were noted in progesterone or DHEA‐S levels at delivery or during the pregnancy; however, estradiol levels were higher (P < 0.05) in the four areas studied at delivery and in the maternal blood with advancing gestation. Additionally, estrone levels tended to increase more rapidly (P = 0.057) in the maternal blood of monkeys receiving genistein than in untreated controls, suggesting that genistein may stimulate the deconjugation of estrone in the gut, thus allowing its reabsorption into the peripheral circulation and conversion to estradiol.

Effect of genistein on steroid hormone production in the pregnant rhesus monkey.

Genistein is a phytoestrogen found in soy beans. Phytoestrogens have been reported to cause reproductive problems in sheep and rats. This research was conducted to determine the effects of genistein fed to rhesus monkeys during pregnancy, with specific interest on fetal growth and steroidogenesis in the maternal-fetoplacental unit. Two groups of five monkeys each were selected in early stages of pregnancy. One group was administered genistein in a fruit treat each weekday until Cesarean section 10 days prior to term. The second, control group, received fruit treats without genistein. Maternal blood samples were collected on Tuesday and Friday of each week. At delivery, samples were collected from the maternal peripheral circulation, uterine veins, uterine-ovarian veins, and the fetal heart. Comparisons between control and genistein-treated monkeys revealed no differences in the maternal weight gained during pregnancy, or in fetal weights or placental weights at delivery. Serum was assayed by radioimmunoassay (RIA) for estradiol, progesterone, dehydroepiandrosterone sulfate (DHEA-S), and estrone. No significant differences (P > 0.05) were noted in progesterone or DHEA-S levels at delivery or during the pregnancy; however, estradiol levels were higher (P < 0.05) in the four areas studied at delivery and in the maternal blood with advancing gestation. Additionally, estrone levels tended to increase more rapidly (P = 0. 057) in the maternal blood of monkeys receiving genistein than in untreated controls, suggesting that genistein may stimulate the deconjugation of estrone in the gut, thus allowing its reabsorption into the peripheral circulation and conversion to estradiol.

JTE-607, a novel inflammatory cytokine synthesis inhibitor without immunosuppression, protects from endotoxin shock in mice.

We investigated the effect of a novel N-benzoyl-L-phenylalanine derivative compound (JTE-607) on production of various cytokines and other immune responses in vitro and on endotoxin shock in vivo. Human, monkey, rabbit, mouse and rat peripheral blood mononuclear cells (PBMCs), and human fibroblasts, umbilical vein endothelial cells (HUVEC), mesangial cells and T cells were used in vitro. Endotoxin shock was induced by lipopolysaccharide (LPS) in Corynebacterium parvum (C. parvum) sensitized male C57BL/6 mice in vivo. JTE-607 inhibited inflammatory cytokine production, including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, IL-8 and IL-10, from LPS-stimulated human PBMCs, with IC50 values of 11, 5.9, 8.8, 7.3 and 9.1 nM, respectively. The inhibitory effects of JTE-607 were also seen in mRNA expression of those cytokines. The potency of JTE-607 on cytokine production from PBMCs of other species, and from other human cells were much lower than that on human PBMCs. JTE-607 did not affect either LPS-stimulated microbead phagocytosis or reactive oxygen species production at 1 microM in human PBMCs but slightly suppressed expression of major histocompatibility complex class II antigen at 1 microM, although it was 100-fold less active than it was as a cytokine inhibitor. JTE-607 (0.3-10 mg/kg, i.v.) showed dose dependent inhibition of mortality after LPS challenge in C. parvum sensitized mice in accordance with a decrease of plasma TNF-alpha. These results suggest that JTE-607 is a multiple cytokine inhibitor specific for human PBMCs. This compound may be useful for the treatment of various cytokine mediated diseases such as septic shock without causing immunosuppression.

The influence of ocular pulsatility on scanning laser Doppler flowmetry

PURPOSE: To determine the effect of the cardiac cycle on scanning laser Doppler flowmeter measurements of retinal capillary blood flow in rhesus monkeys and humans.METHODS: Multiple scanning laser Doppler flowmetry images of rhesus monkey and human retinal capillary blood flow over a range of heart rates were obtained. Average flow values were determined for the 64 scan lines that compose the two-dimensional flow map. Cutaneous blood flow was measured simultaneously with a laser Doppler flowmeter. The temporal relationships between retinal capillary blood flow, peripheral arterial pulse, and cutaneous blood flow were determined. In addition, human retinal capillary blood flow in a 10 × 10-pixel area during different phases of the cardiac cycle was compared.RESULTS: Regular oscillations in human and rhesus monkey retinal capillary blood flow are evident as alternating bright and dark horizontal bands in scanning laser Doppler flowmetry images. These fluctuations are temporally correlated with cutaneous blood flow. Linear regression of actual vs predicted heart rate based on peaks in retinal capillary flow yielded r = 0.999 in a rhesus monkey and 0.938 in a human. Retinal capillary blood flow in a 10 × 10-pixel area fluctuated as much as 50% depending on the phase of the cardiac cycle.CONCLUSIONS: The alternating bright and dark banding pattern observed in scanning laser Doppler flowmetry scans of retinal capillary blood flow is related to the cardiac pulse. The errors introduced by pulse-related fluctuations in retinal capillary blood flow are significant and must be minimized or corrected for accurate and reproducible measurements of ocular hemodynamics.

Long-term discordant xenogeneic (porcine-to-primate) bone marrow engraftment in a monkey treated with porcine-specific growth factors.

Mixed allogeneic hematopoietic chimerism has previously been reliably achieved and shown to induce tolerance to fully MHC-mismatched allografts in mice and monkeys. However, the establishment of hematopoietic chimerism has been difficult to achieve in the discordant pig-to-primate xenogeneic model. To address this issue, two cynomolgus monkeys were conditioned by whole body irradiation (total dose 300 cGy) 6 and 5 days before the infusion of pig bone marrow (BM). Monkey anti-pig natural antibodies were immunoadsorbed by extracorporeal perfusion of monkey blood through a pig liver, immediately before the intravenous infusion of porcine BM (day 0). Cyclosporine was administered for 4 weeks and 15-deoxyspergualin for 2 weeks. One monkey received recombinant pig cytokines (stem cell factor and interleukin 3) for 2 weeks, whereas the other received only saline as a control. Both monkeys recovered from pancytopenia within 4 weeks of whole body irradiation. Anti-pig IgM and IgG antibodies were successfully depleted by the liver perfusion but returned to pretreatment levels within 12-14 days. Methylcellulose colony assays at days 180 and 300 revealed that about 2% of the myeloid progenitors in the BM of the cytokine-treated recipient were of pig origin, whereas no chimerism was detected in the BM of the untreated control monkey at similar times. The chimeric animal was less responsive by mixed lymphocyte reaction to pig-specific stimulators than the control monkey and significantly hyporesponsive when compared with a monkey that had rejected a porcine kidney transplant. To our knowledge, this is the first report of long-term survival of discordant xenogeneic BM in a primate recipient.

Behavioral Impairment Produced by Low-Level Postnatal PCB Exposure in Monkeys

The preponderance of evidence in humans suggests that polychlorinated biphenyl (PCB)-induced behavioral deficits result from prenatal exposure rather than exposure through breast milk, although a recent study reported lower psychomotor scores during infancy associated with PCB concentration in breast milk. In the current study, monkeys were dosed from birth to 20 weeks of age with a PCB congener mixture representative of the PCBs found in human breast milk. Blood and fat levels of PCB-exposed monkeys at the end of the dosing period were within the range observed in the general human population, while levels in control monkeys were below averages observed in humans in industrialized countries. Behavioral assessment on a series of tasks was performed when monkeys were between 2.5 and 5.0 years of age. Robust deficits were observed on spatial delayed alternation, fixed interval, and differential reinforcement of low rate performance. No group differences were observed for the number of errors on a series of nonspatial and spatial discrimination reversal tasks. Behavioral deficits included retarded learning, perseverative behavior, and inability to inhibit inappropriate responding. These results have implications for the potential contribution of exposure to PCBs through breast milk to behavioral impairment.

Ability of teicoplanin and vancomycin to induce contraction of, and histamine release from, pulmonary tissue of humans, monkeys and guinea pigs

To assess the safety of teicoplanin and vancomycin with respect to airway tissue, we evaluated whether these two antibiotics induce pulmonary tissue contraction and histamine release in human, monkey and guinea pig specimens in vitro. The effects of these drugs on the release of histamine from monkey blood leucocytes and mouse bone marrow-derived mast cells (BMMC) were also studied. Neither teicoplanin nor vancomycin (10(-6)-10(-3) g/mL) induced contractions of guinea pig trachea or lung parenchyma. Similarly, these drugs induced no appreciable change in the resting tonus of cynomolgus monkey bronchus or lung parenchyma. The tonus of monkey trachea was not influenced by teicoplanin, whereas 10(-3) g/mL vancomycin caused contraction. The spontaneous tonus of human lung parenchyma was not altered by teicoplanin or vancomycin, and that of the bronchus was not influenced by teicoplanin; however, 10(-3) g/mL vancomycin elicited obvious contraction of the bronchus. Neither drug promoted the release of significant amounts of histamine from these pulmonary tissues or from monkey blood leucocytes and BMMC. These results suggest that, compared with vancomycin, teicoplanin may be associated with a lower risk of inducing bronchospasm when used for inhalation therapy.

Effects of space flight on surface marker expression

Space flight has been shown to affect expression of several cell surface markers. These markers play important roles in regulation of immune responses, including CD4 and CD8. The studies have involved flight of experimental animals and humans followed by analysis of tissue samples (blood in humans, rats and monkeys, spleen, thymus, lymph nodes and bone marrow in rodents). The degree and direction of the changes induced by space flight have been determined by the conditions of the flight. Also, there may be compartmentalization of the response of surface markers to space flight, with differences in the response of cells isolated from blood and local immune tissue. The same type of compartmentalization was also observed with cell adhesion molecules (integrins). In this case, the expression of integrins from lymph node cells differed from that of splenocytes isolated from rats immediately after space flight. Cell culture studies have indicated that there may be an inhibition in conversion of a precursor cell line to cells exhibiting mature macrophage characteristics after space flight, however, these experiments were limited as a result of technical difficulties. In general, it is clear that space flight results in alterations of cell surface markers. The biological significance of these changes remains to be established.

Mosquitoes Mating on the Metro

Chen Xiao Ping from China asked some curious questions about such infections, including what complications occur, what dose induces infection without complications, how the immune response compares with that of P. vivax, what other infections would be caused by injection of monkey blood into people, and ‘How about the ethics of clinical trial using P. cynomolgi in human bodies?’ Ole Skovmand (ORSTOM, Montpellier, France) assumed the questions were academic, and pointed out that monkeys share many infectious diseases with people and that HIV in particular may have been of simian origin. Kevin Baird (Naval Medical Research Institute, Maryland, USA) said that an excellent history of P. cynomolgi infection of human beings can be found in the classic treatise ‘The Primate Malarias’ by G.R. Coatney et al., printed by the US Department of Health, Education and Welfare (now the Department of Health & Human Services), National Institutes of Health (1971). ‘It would be wonderful if Nih took the trouble and expense to reprint this book…P. cynomolgi can infect humans by mosquito bite [which] eliminates the risks…that would attend challenge with blood stages.’ Whether the experiment is ethical depends upon objectives and its safety, and how these are reviewed by an ethical board.

Quantitative analysis of acetaldehyde in whole blood from human and various animals by gas chromatography

Acetaldehyde present in the blood of bull, chicken, hamster, horse, human, monkey, pig, rabbit, rat and sheep, was quantitatively analyzed by a newly developed gas chromatographic method. Acetaldehyde in a blood sample was reacted with cysteamine to give 2-methylthiazolidine, which was extracted with dichloromethane and subsequently analyzed by gas chromatography with a fused-silica capillary column and a nitrogen–phosphorus detector. The quantities of acetaldehyde found in blood ranged from 2.04 μmol/ml (hamster) to 14.8 μmol/ml (pig). The quantity of acetaldehyde recovered from human blood was 6.17 μmol/ml.

5' nuclease PCR assay to detect Yersinia pestis.

The 5' nuclease PCR assay uses a fluorescently labeled oligonucleotide probe (TaqMan) to rapidly detect and quantitate DNA templates in clinical samples. We developed a 5' nuclease PCR assay targeting the plasminogen activator gene (pla) of Yersinia pestis. The assay is species specific, with a detection threshold of 2.1 x 10(5) copies of the pla target or 1.6 pg of total cell DNA. The assay detected Y. pestis in experimentally infected Xenopsylla cheopis fleas and in experimentally infected monkey blood and oropharyngeal swabs. The TaqMan assay is simple to perform and rapid and shows promise as a future field-adaptable technique.

Spectrophotometric and Spectrofluorimetric Methods for the Determination of Terazosin in Dosage Forms

Five simple and accurate methods are presented for the determination of terazosin (TZ) in tablets. These methods are based on: the direct measurements of the first and second derivative spectra of samples (A), the reaction of TZ with chloranil (CH) in aqueous solution of pH 9 to give an intense yellow color measured at 340 nm (B), the reaction of the drug with mercurochrome (MER) in aqueous alkaline medium to give an intense red color measured at 543 nm (C), the formation of an ion-pair salt between the drug and bromocresol purple (BCP) with subsequent absorbance measurements at 412 nm (D), and a sensitive fluorimetric method (E). The latter method was extended to determine TZ in presence of its degradation products.

Removal of primate xenoreactive natural antibodies by extracorporeal perfusion of pig kidneys and livers

Organ perfusion is one of the possible strategies to attenuate rejection of discordant xenografts by reducing the levels of the recipient's xenoreactive natural antibodies (XNA). Its efficacy in terms of XNA removal was studied in models of primate blood or plasma perfusion through porcine-kidneys or livers, which special attention to haematology consequences and potential side-effects. We first perfused the blood of rhesus monkeys through pig kidneys and livers, and demonstrated that the perfusion of a pig liver resulted in higher XNA adsorption (72 ± 13%) than the perfusion of a pig kidney (51 ± 25%). However, when we normalized for the weight of the perfused organs and for levels of natural antibodies in individual monkeys, livers adsorbed less antibody (1.4 ± 0.9 U antibody/g) than kidneys (7.2 ± 7 U antibody/g). Histological signs of rejection were observed in perfused kidneys, but not in perfused livers. A major drawback of the perfusion of blood through livers was a considerable decrease in the primates' haemoglobin and platelet levels. To avoid this, we developed a plasma liver perfusion device. This method allowed a significant improvement in the haemodynamic states of primates and was particularly effective in preventing anaemia. Moreover, plasma liver perfusion was as effective as blood liver perfusion to remove natural antibodies and, resulted in a marked decrease in their functional activity as assessed by complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC). The level of other plasma proteins was not significantly affected, apart from a dilution effect. After xenoperfusion a strong antibody response was evidenced by ELISA, CDC and ADCC between days 7 and 14 and then decreased progressively. We conclude that the separation of blood to allow the perfusion of plasma through a pig organ is safer than the perfusion of unseparated blood and is associated with efficient natural antibody removal. However, organ perfusion is limited by a rebound in antibody levels after a few days, and thus will have to be associated with anti-B cell immunosuppressive therapy for long-term or repeated applications.