Mixed haemadsorption should be regarded as an application of the mixed antiglobulin reaction to situations where the antigen is sessile on a glass surface. Antibody attached to the antigen when exposing the latter to an antiserum is traced by red cells carrying an antiglobulin layer which makes them adsorb to the antibody.
The indicator cells are prepared by coating them first with a layer of γ-globulin from the animal species, the antibody globulin of which they are intended to trace, and then with a layer of the corresponding antiglobulin. The most effective indicator cells were obtained by attaching antibody to natural receptors on the red cells to achieve their first coating of γ-globulin.
The preparation of indicator cells for tracing antibodies from a number of species, including human, is described.
The mixed haemadsorption technique is highly specific and has a sensitivity which is comparable to that of the most sensitive serological techniques.
Test procedures adapted for different purposes are outlined and a number of applications to experimental and clinical problems are reviewed.
The occlusion of vessels by packed Plasmodium falciparum-infected (iRBC) and uninfected erythrocytes is a characteristic postmortem finding in the microvasculature of patients with severe malaria. Here we have employed immunocompetent Sprague-Dawley rats to establish sequestration in vivo. Human iRBC cultivated in vitro and purified in a single step over a magnet were labeled with 99mtechnetium, injected into the tail vein of the rat, and monitored dynamically for adhesion in the microvasculature using whole-body imaging or imaging of the lungs subsequent to surgical removal. iRBC of different lines and clones sequester avidly in vivo while uninfected erythrocytes did not. Histological examination revealed that a multiadhesive parasite adhered in the larger microvasculature, inducing extensive intravascular changes while CD36- and chondroitin sulfate A-specific parasites predominantly sequester in capillaries, inducing no or minor pathology. Removal of the adhesive ligand Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), preincubation of the iRBC with sera to PfEMP1 or preincubation with soluble PfEMP1-receptors prior to injection significantly reduced the sequestration. The specificity of iRBC binding to the heterologous murine receptors was confirmed in vitro, using primary rat lung endothelial cells and rat lung cryosections. In offering flow dynamics, nonmanipulated endothelial cells, and an intact immune system, we believe this syngeneic animal model to be an important complement to existing in vitro systems for the screening of vaccines and adjunct therapies aiming at the prevention and treatment of severe malaria. 相似文献
We have studied the location and mechanism of apoptosis within the granulomas in the lungs at various stages of slowly progressive primary murine Mycobacterium tuberculosis infection. Parallel sections were analyzed for detection of mycobacterial antigens, Fas, and Fas ligand (FasL) by immunohistochemistry, and for apoptotic cells by terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) method. The frequency of apoptosis was high in the macrophage aggregates as compared to the lymphocyte aggregates and at the interface between them. Five to seven percent of the vacuolated macrophages in the granulomas expressed FasL intensely. These cells contained large amounts of mycobacterial antigens. These findings suggest that M. tuberculosis infection can induce increased expression of FasL in a population of infected macrophages. As a consequence the infected macrophages will be protected from the attack of cytotoxic T cells and activation of bactericidal mechanisms by Th1 type lymphocytes. This constitutes a novel evasion mechanism for M. tuberculosis possibly explaining the chronic course of infection. 相似文献
Five gram-positive bacterial strains were selected for absorption studies of human serum samples. Strain AR1 (group A, M-type 1) and G148 (group G), with strong immunoglobulin G (IgG) binding capacities, and strain AW43 (group A, M-type 60), binding both IgA1 and IgA2, were compared with Staphylococcus aureus Cowan I and with Staphylococcus epidermidis L603. Both AR1 and G148 were capable of completely absorbing out serum IgG. In contrast, S. aureus Cowan I left a fraction unabsorbed, as expected from its known lack of IgG3 binding. Strain AW43 absorbed out all serum IgA, using a 10-microliter bacterial pellet for 20 microliter of serum. Serum IgM levels were slightly reduced by S. aureus Cowan I absorption. On the basis of the experiments, a bacterial mixture was designed consisting of S. aureus Cowan I and group A streptococcus strains AR1 and AW43, with absorption characteristics suitable for use in discriminating between early IgM and late IgG and IgA immune responses in routine serological work. A new type of bacteria-mammalian protein binding was discovered. Human serum albumin was completely absorbed out by strain G148 and to a lesser extent by strain AR1 and AW43. S. aureus Cowan I and S. epidermidis were negative. The binding capacity of G148 for albumin equalled that of Cowan I for IgG. The binding pattern of albumin to the strains was different from those of IgG, IgA, IgM, fibrinogen, haptoglobin, or aggregated beta 2-microglobulin and therefore seems to represent another type of bacterial-mammalian interaction with a specific albumin receptor on the surface of streptococci. 相似文献
We have studied the relationships between genital or rectal carriage of group B streptococci (GBS) with the levels of systemic and mucosal antibodies to GBS in 200 women at about week 17 of pregnancy. Secretions from the uterine cervix were collected with absorbent cylindrical wicks for quantification of antibody levels with whole cell enzyme-linked immunosorbent assay. GBS were cultured from the cervix (with or without concomitant rectal colonization) of 13.5%, from the rectum (with or without concomitant cervical colonization) of 12%, and from both culture sites of 8.5% of the women. Serotypes Ia, II, and III were predominant. Compared with culture-negative women, the group of women colonized rectally had markedly elevated levels of both immunoglobulin A (IgA) and IgG antibodies to GBS in cervical secretions and also had a moderate but significant elevation of IgA antibodies in sera. Women colonized only in the cervix had increases of specific IgA and IgG antibodies in cervical secretions, but their serum antibody levels were not elevated. In cervical secretions, the increase in antibody levels in the groups of colonized women was most pronounced for the IgG isotype, indicating a mucosal immune response involving IgG as well as IgA. A close correlation was found among the levels of antibodies to each of the three GBS serotypes tested. Evidence for such cross-reacting antibodies to different serotypes of GBS, as well as to group A streptococci, was also obtained from absorption experiments. Altogether, our results show that undiluted secretions for antibody determination can be easily collected from the uterine cervix with absorbent wicks and demonstrate that colonization of GBS in the rectum and the uterine cervix may induce a systemic as well as a pronounced local immune response in the female genital tract. The findings may have implications for the development of a mucosal vaccine against GBS disease. 相似文献
Extracellular superoxide dismutase (EC-SOD) controls the availability of extracellular superoxide (O
2-
), which is important for a variety of physiological pathways, including the primary means of inactivating nitric oxide (NO). The role of EC-SOD in neurobehavioral function has been until now unexplored. In the current studies, the phenotypic expression of genotypic alterations of EC-SOD production in mice were characterized for spatial learning and memory. Dramatic impairments in spatial learning in the win-shift 8-arm radial maze were seen in both EC-SOD knockout mice and EC-SOD overexpressing mice. The EC-SOD overexpressing mice were further characterized as having significant deficits in a repeated acquisition task in the radial-arm maze, which permitted the dissociation of long and short-term learning. Long-term learning was significantly impaired by EC-SOD overexpression, whereas short-term learning was not significantly affected by EC-SOD overexpression. NO systems have been shown to be importantly involved in learning and memory. This may be important in the current studies because EC-SOD has primary control over the inactivation of NO. We found that EC-SOD overexpressing mice were resistant to the cognitive effects of L-NAME (NG-nitro-L-arginine methyl ester hydrochloride), an NO synthase inhibitor. Decreased NO catabolism in these mice may have served to counter the effects of NOS inhibition by L-NAME. The current finding that EC-SOD levels that were either higher or lower than controls impaired learning demonstrates that the proper control of brain extracellular (O
2-
) may be more vital than merely reduction of brain extracelluar (O
2-
) in maintaining adequate learning function. 相似文献
Summary Intrathecal administration of the substance P antagonist Spantide caused marked necrotic changes of the gray matter of the spinal cord extending several segments from the injection site. Intravenous treatment with several doses of thyrotropin releasing hormone before and after Spantide injection completely prevented the necrotic lesion. 相似文献
The nosologic status of fibrous dysplasia (FD), a well-known and relatively common bone lesion, is controversial. Information collected by the CHromosomes And MorPhology (CHAMP) study group on published and unpublished cases of fibrous dysplasia shows the presence of clonal chromosome changes in at least a proportion of these lesions. The chromosome aberrations found in FD lesions have been quite variable and have included both structural and numerical changes. Two of the three cases investigated at the study group had trisomy 2 as the sole acquired anomaly. Combined with previously published data, +2 and rearrangements involving chromosome band 12p13 have each been detected in 3 of 8 cases with abnormal karyotype of 11 in which chromosomal analysis has been performed, suggesting that FD is a neoplastic lesion rather than a "dysplastic" process, as has been generally believed and as implied by its very name. 相似文献
Pressure sores are a current problem in hospitals and care of the elderly, leading to protracted hospital stays and a high
care burden. The trauma for the patients is severe, and the cost of pressure sore prevention and treatment, is considerable.
Antidecubitus mattresses are used for prevention and in treatment, but they also contribute to the cost of treating pressure
sores. The problem highlighted in the review is that the mattresses' effectiveness in preventing and treating pressure sores
has not been sufficiently evaluated. When antidecubitus mattresses are evaluated, it is often only with regard to aspects
of the interface pressure and the mattresses' ability to redistribute the pressure. The review points out the important observation
that, to be able to evaluate the efficacy of the antidecubitus mattress, the mattress's effect on tissue viability needs to
be studied. The parameters that ought to be considered when evaluating a support surface are: interface pressure, pressure
and blood flow distribution, temperature and humidity in the skin-support surface interface. The authors propose that the
effect on tissue viability of external loading can be assessed by simultaneous measurement of the interface pressure and tissue
perfusion. 相似文献