Use of the collagen-binding assay for von Willebrand factor in the analysis of type 2M von Willebrand disease: a comparison with the ristocetin cofactor assay

This study compares the utility of two functional assays for von Willebrand factor (VWF), the ristocetin cofactor assay (VWF:RCo) and the collagen-binding assay (VWF:CBA). We analysed a group of 32 patients with type 2 von Willebrand disease (VWD) (25 patients with type 2M, six with type 2A and one with type 2B) and 22 normal control subjects. VWF:RCo/VWF antigen (VWF:Ag) ratios and VWF:CBA/VWF:Ag ratios were compared between the patient and control groups. In the six patients with type 2A VWD, both VWF:RCo/VWF:Ag ratios and VWF:CBA/VWF:Ag ratios were discordant (< or = 0.7). In the 25 type 2M VWD patients, the VWF:CBA/VWF:Ag ratios were concordant (> 0.7), but the VWF:RCo/VWF:CBA ratios were discordant (< or = 0.7) (P = 0.001) compared with control subjects. Thus, VWF:RCo/VWF:Ag ratios were discordant in both type 2M and 2A VWD patient groups indicating a functional abnormality. However, VWF:CBA/VWF:Ag ratios were discordant in the type 2A VWD group but not in the type 2M VWD group. Our study showed that VWF:CBA is sensitive to functional variants associated with the loss of high-molecular-weight multimers, i.e. type 2A and 2B in VWD, but the assay was unable to discriminate defective platelet-binding VWD variants with normal multimeric patterns such as type 2M VWD. It was concluded that the VWF:CBA assay should be used in association with rather than as a replacement for the VWF:RCo assay.     

Schistosoma mansoni: histological analysis of the synergistic interaction between vaccine immunity and praziquantel therapy in the lungs of mice

Summary Naive CBA mice and mice vaccinated 4 weeks previously with gamma-irradiated cercariae of 5. mansoni were challenged percutaneously with normal cercariae and then treated with 500 mg/kg body weight of Praziquantel (Pzq). The drug was administered intradermally on day 1 or intramuscularly on day 6, thus targeting against skin stage or lung stage challenge larvae respectively. The skin site of challenge and/or the lungs were removed at various time points to provide samples for histological examination. As reported elsewhere (Flisser, Delgado & McLaren 1989) the efficacy of Pzq was significantly enhanced in vaccinated mice and was influenced by the treatment regime. Histological analysis revealed that when Pzq was administered I/D on day 1 to vaccinated mice, the inflammatory response to challenge differed in extent but not nature from that seen in vaccinated but untreated cohorts. This correlates with worm recovery data showing no (this study), or only marginal synergy between drug treatment and immunity using this regimen of drug treatment (Flisser et al. 1989). Following the day 6 protocol of drug delivery, however, lungs from treated vaccinated mice exhibited many large inflammatory reactions containing trapped challenge larvae. In contrast, lungs from untreated vaccinated mice had only few foci which were small and rarely contained trapped larvae. These data again correlate well with worm recovery data showing that there is a highly significant synergy between vaccination and drug treatment administered at this time (Flisser et al. 1989; this study). It would seem, therefore, that Pzq exacerbates lung phase immunity in the NIMR vaccine mouse model where skin phase immunity predominates and pulmonary attrition is normally minimal. The results are discussed in the light of published data concerning the effector mechanisms thought to characterize skin and lung phase vaccine resistance in the murine model.     

Kinetics of immunoglobulin and specific antibody responses of CBA mice infected with Trypanosoma rhodesiense

Summary Groups of CBA/CaJ and B-cell deficient CBA/N mice were infected with Trypanosoma rhodesiense EATRO 1886 strain. Survival, parasitaemia, serum Ig levels plus specific trypanosomal IgM and IgG antibodies were assayed and compared during infection. Whereas both strains of mice had similar parasitaemias during the first week of infection, CBA/N parasitaemias were lower than those observed in CBA/CaJ mice during the subsequent study period. Antibody responses, specific for T. rhodesiense antigens, peaked on day 10 after infection in CBA/CaJ mice, then rapidly declined. However, antibody responses in CBA/N mice remained elevated throughout the study. In addition, the kinetics of specific IgG and IgM varied in CBA/N mice: IgG antibody was detected on day 4, whereas specific IgM was detected on day 16. This unique relationship between the appearance of IgG and IgM antibody may explain the longer survival observed for B-cell deficient CBA/N mice infected with T. rhodesiense.     

Effect of smoking on immunity in human chronic periodontitis

Aim

Evaluate the effects of smoking on dendritic cells (DCs), cytokines, clinical periodontal parameters, and number of teeth in samples of human chronic periodontitis (CP).

Material and methods

Gingival samples were obtained from 24 smokers and 21 non-smokers with CP. Periodontal examination was carried out. Immunohistochemical staining was performed to identify Factor XIIIa+ immature, CD1a+ immature, and CD83+ mature DCs. The inflammatory infiltrate was counted, and IL-2, IL-10, IL-4, IL-6, IFN-γ, TNF-α, and IL-17A were measured using the cytometric bead array (CBA). Inflammatory infiltrate, DCs, cytokines, classification of CP, clinical periodontal parameters, number of teeth, smoking habit in years (SH/years), and number of cigarettes smoked per day (C/day) were correlated and compared.

Results

CD83+ mature DCs decreased in the smokers group. Negative correlations could be observed between the number of C/day with levels of IL-17A and number of teeth. Correlations between smoking, periodontal disease status, and other cytokines were not observed.

Conclusions

Smoking decreases mature DCs in chronic periodontitis. Moreover, a dose-dependent relation can be observed between C/day and number of teeth and levels of IL17A observed. Smokers show a different modulation of the CP immune response.     

Sudden outbreak of a leukemia-like lesion in female CBA mice after repeated injections of 5-azacytidine

Summary A total of 11 i.p. injections of 1 mg/kg 5-azacytidine given within 21 weeks induced a sudden outbreak of a leukemia-like disease in female CBA mice about 2 weeks after the last injection. The outbreak of disease was highly synchronous; 68% of animals who had survived the treatment period succumbed within a period of 5 days, 2/3 of them on a single day. The lesion had a characteristic form commonly presenting as a result of hemorrhagic effusion in the thoracic and/or abdominal cavities, associated with diffuse infiltration of lymphatic and fatty tissue by lymphoblasts. It appears that a critical combination of factors was responsible for the unexpected appearance of the lesion. Female BALB/c mice treated identically were not affected. The factors indicating that the lesion was induced by an epigenetic mechanism are discussed.In association with EURATOM, Contract No. BI6-0080-D(B)     

Swimming navigation,open-field activity,and extrapolation behavior of two inbred mouse strains with Robertsonian translocation of chromosomes 8 and 17

Female mice from inbred strains carrying a Robertsonian translocation (nine CBARb and eight C57BL/6Rb) were compared with animals from their respective strains (seven CBA and nine C57BL/6) first in open-field activity (two exposures of 10-min duration), then during 5 days (with six trials each) in Morris' swimming navigation test, and finally, in their ability to extrapolate the future position of a food reward being moved slowly out of their reach. ANOVA (strain and translocation) revealed significant effects of Robertsonian translocations (Rb) in swimming navigation,Rb mice being impaired primarily in the initial phases of acquisition and during the first trials of platform reversal and the impairment being stronger in C57BL/6 mice. In the open field,Rb mice were as active as the normal strains but showed significantly increased path tortuosity and moved slightly faster. In the extrapolation task,Rb mice showed above-chance levels in moving to the target indicated by the disappearance of the stimulus, while normal mice chose at chance levels, but the translocation effects were not statistically significant. These data indicate that telocentric fusion of chromosomes may entail behavioral alterations, perhaps by subtle changes in neurotransmitters or limbic circuitry. The expression of such alterations, however, can be remarkably strain dependent.     

Selective expansion of idiotype sharing T and B cells in cyclosporin A-mediated autoimmunity.

CBA/N mice submitted to autologous bone marrow reconstitution after lethal irradiation and simultaneous Cyclosporin A (CsA) treatment develop a chronic graft-versus-host disease with autoimmune characteristics. When compared to normal controls, diseased mice show an overrepresentation of V beta 8-expressing T cells (65-80% of all CD3+ lymphocytes), together with a marked increase in the titres of serum Ig that specifically bind to F(ab')2 fragments of anti-V beta 8 F23.1 antibodies. Such 'V beta 8-like' Ig V regions are abundantly represented among the IgG2b and mAbs of an unselected collection of hybridomas derived from these mice. These mAbs are not multireactive Ig as they fail to bind to a panel of various antigens and antibodies, but often show simultaneous reactivity with anti-idiotypic mAbs to F23.1 and auto-binding. These molecules may provide the structural basis of V-region specific complementarities, driving the expansion of restricted T and B cell repertoires associated with pathological autoimmunity.     

Differences in the training capacity of 101/H and CBA mice in a water labyrinth (Modified Morris' test)

The rate of formation of the habit of search for submerged platform was studied by the modified Morris' test in 101/H and CBA mice. The former strain was trained slower than the latter; in addition, 101/H mice of different sexes differed by the time of latent periods of searching for the platform. The results are interpreted as a manifestation of genetically determined disorders of training and/or memory in 101/H mice. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 12, pp. 666–668, December, 1997