Absence of Correlation Between Delayed-Type Hypersensitivity and Protection in Experimental Systemic Candidiasis in Immunized Mice

We found that in mice which had been immunized intraperitoneally with 2 × 10⁸ heat-killed Candida albicans cells there was a striking temporal relationship between resistance to systemic challenge with 10⁶ living C. albicans cells and a number of measurable cellular parameters of the host response. These included the emergence of delayed-type hypersensitivity and the development of granulocytosis. Since it had been shown in previous work that granulocytosis was associated with an increase in resistance when nonspecific immunostimulation was used, we performed experiments to induce delayed-type hypersensitivity without any measurable modification of the granulocyte population. Adoptive transfer of delayed-type hypersensitivity with spleen cells from immune and resistant donor mice did not produce any increase in resistance in normal recipients. When separate groups of mice were immunized intraperitoneally or subcutaneously with varying doses of heat-killed C. albicans, we found that doses of less than 10⁸ cells did induce significant delayed-type hypersensitivity without any increase in granulocytosis. In such mice, as well as in animals pretreated with immunomodulators before immunization with heat-killed C. albicans, the presence of cell-mediated immunity, as measured by the delayed-type hypersensitivity test, was not associated with an increase in resistance against systemic candidiasis. On the contrary, the results suggested that cell-mediated immunity was associated with an increase in the susceptibility of these mice. The same effect on candidiasis susceptibility was observed when animals were immunized with heat-killed filamentous C. albicans.     

Expression of high- and low-affinity receptors for C3a on the human mast cell line,HMC-1

The proteolytic cleavage product of complement component 3, (C3a), like C4a and C5a, is a potent anaphylatoxin and induces the production of inflammatory mediators in phagocytes. Notably, mast cells respond to C3a with the release of vasoactive substances, including histamine. We have examined the function and receptor binding of C3a in a human leukemic mast cell line, HMC-1. Similar to chemoattractant agonists in leukocytes, C3a induced rapid cytosolic free calcium concentration increases in HMC-1 cells. EGTA did not diminish this response, indicating that mobilizable Ca²⁺ was from intracellular stores. Receptors for C3a in HMC-1 cells couple in part to Bordetella pertussis toxin-sensitive G-proteins and, therefore, appear to belong to the family of serpentine receptors that require G-proteins for signal transduction. HMC-1 cells express two types of C3a receptors, C3aR1 and C3aR2, that were shown to bind ¹²⁵I-C3a with high-(K_d1 = 2.1–4.8 nM) or low-affinity (K_d2 = 30–150 nM), and both receptors are expressed at high level: 3 × 10⁵–6 × 10⁵ C3aR1/cell and 5 × 10⁵–2.3 × 10⁶ C3aR2/cell. Results from cross-linking experiments with ¹²⁵I-C3a fully agree with the presence of two different classes of C3a receptors in HMC-1 cells. Two membrane proteins with apparent molecular masses of 54–61 kDa (p57) and 86–107 kDa (p97) could be covalently modified with ¹²⁵I-C3a, and this cross-linking was inhibited with an excess of unlabeled C3a. Many of the known agonists for leukocytes including 13 chemokines (IL-8, NAP-2, GROα, ENA-78, IP10, PF4, MCP-1, 2 and 3, RANTES, MIP-1α, MIP-1β and 1309), three neuropeptides (neuropeptide Y, somatostatin and calcitonin), as well as C5a, did not activate HMC-1 cells, indicating that C3a is one of a few protein ligands for which this cell line expresses specific receptors. The apparent selectivity for C3a and the abundant expression of C3a receptors make the HMC-1 cell line an excellent choice for the cloning of the receptor genes.     

Interleukin-10 inhibits IgE-mediated nitric oxide synthase induction and cytokine synthesis in normal human keratinocytes

Human keratinocytes (HK) generate nitric oxide (NO) and proinflammatory mediators following activation with either IgE/anti-IgE immune complexes or a combination of lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Recently, interleukin-10 (IL-10) has been shown to down-regulate various inflammatory responses and to be secreted by lymphocytes and dendritic cells during skin inflammatory reactions. We show here that IL-10 down-regulates the production of tumor necrosis factor (TNF)-α and IL-6 by activated HK. Also, induction of inducible nitric oxide synthase (iNOS) expression in HK by IgE/anti-IgE or LPS/IFN-γ is significantly reduced by the addition of IL-10. This effect is dose dependent and correlates with reduction of iNOS mRNA production and enzyme level. Therefore, IL-10 down-regulates NO-mediated HK inflammatory responses and may thus participate in the regulation of the skin immune network.     

Immunogenicity and protective efficacy of the alpha C protein of group B streptococci are inversely related to the number of repeats.

Infection by group B streptococci (GBS) is an important cause of bacterial disease in neonates. Alpha C protein is a protective cell surface-associated protein of GBS. This protein contains a repeat region flanked by N and C termini. Variable expression of tandem repeating units of alpha C proteins had been found among clinical isolates of GBS. We examined the effect of the number of repeats on the immunogenicity of the alpha C protein and its ability to elicit protection from GBS infection in a neonatal mouse model. Mice were immunized with purified alpha C proteins of constructs containing various numbers of repeats (n = 1, 2, 9, and 16) and the N- and C-terminal regions. Both the N-terminal and the repeat regions contain protective and opsonic epitopes. Antibody responses to the alpha C protein constructs with various numbers of repeats were tested with enzyme-linked immunosorbent assay plates coated with either native, nine-repeat alpha C protein or "repeatless" N-terminal antigen. An inverse relationship was found between the number of repeats and the immunogenicity of the alpha C protein; this effect was most pronounced on titers of antibody to the N-terminal region. An inverse relationship was also observed between the number of repeats and protective efficacy, i.e., mouse dams immunized with 5 microg of one- or nine-repeat alpha C protein transferred protective immunity to 65 or 11% of their pups, respectively (P < 0.0001). Thus, the presence of multiple repeats appears to lessen the antibody response to the complete alpha C protein, and especially the antibody response to its N-terminal region, and suggests a mechanism whereby repeat elements contribute to the evasion of host immunity.     

Computed tomographic study of the posterior condylar angle in arthritic knees: its use in the rotational positioning of the femoral implant of total knee prostheses

The epicondylar axis is a reliable reference to check the rotation of the femoral implant in total knee prostheses (TKPs). However, during the operation it seems easier to use the posterior condylar axis as a landmark. The angle between these two axes is called the posterior condylar angle (PCA). The aim of this study was to measure the PCA in arthritic knees to assess the reliability of the posterior condylar axis as a reference for the control of the rotation of the femoral implant and to look for correlation with other radiological measurements. This prospective study consisted of 103 arthritic knees (81 varus, 22 valgus) before a TKP had been done in 103 patients (75 women, 28 men). The assessment of the PCA was made by computed tomographic scanning (CT). The HKA, HKS and HKT angles were measured on the pangonogram. The posterior condylar axis was internally rotated with respect to the epicondylar axis. The average value for all the patients was 2.65° degrees with a range from 0° to 7°. The PCA was significantly increased in the valgus knees. There was no correlation between the angles on the pangonogram and the posterior condylar axis. While the preoperative assessment of the PCA by CT scanning is reliable, the results obtained indicate the marked variability in its value. If one wishes to use the posterior condylar axis as a guide for rotation, it is therefore necessary to assess the PCA for each patient using adjustable jigs according to the value obtained. No measurement on standard radiographs allowed an extrapolation of the value of the PCA, and CT scanning seems to be the preferable radiological examination.

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Etude tomodensitométrique de l'angle condylien postérieur dans les genoux arthrosiques. Intérêt dans le positionnement en rotation de l'implant fémoral dans les prothèses totales de genou

Résumé L'axe épicondylien est une référence fiable pour le contrôle de la rotation de l'implant fémoral dans les prothèses totales de genou (PTG). Mais, lors de l'intervention, il semble plus facile d'utiliser l'axe condylien postérieur comme repère. L'angle entre ses deux axes est appelé angle condylien postérieur (ACP). Le but de cette étude était de mesurer l'ACP dans les genoux arthrosiques, d'évaluer la fiabilité de l'axe condylien postérieur comme référence pour le réglage de la rotation de l'implant fémoral, de rechercher une corrélation avec d'autres mesures radiologiques. Une étude prospective comportant 103 genoux arthrosiques (81 varus et 22 valgus), avant PTG a été effectuée, chez 103 patients (75 femmes et 28 hommes). L'évaluation de l'ACP a été faite par examen tomodensitométrique (TDM). Les angles HKA, HKS et HKT ont été mesurés sur le pangonogramme. L'axe condylien postérieur était en rotation interne par rapport à l'axe épicondylien. La valeur moyenne pour tous les patients était de 2.65°, avec des valeurs de 0 à 7°. La valeur de l'angle CP augmentait avec une différence significative dans le groupe des genu valgum. Il n'y avait pas de corrélation entre les angles du pangonogramme et l'ACP. Si l'évaluation pré-opératoire de l'ACP par TDM est fiable, les résultats obtenus mettent en évidence une variabilité importante de sa valeur. Il faut donc, si l'on veut utiliser l'axe condylien postérieur comme repère de rotation, évaluer pour chaque patient l'ACP, et utiliser un ancillaire réglable reportant la valeur obtenue. Aucune mesure sur des radiographies standard ne permettant d'extrapoler la valeur de l'ACP, la TDM semble l'examen radiologique de choix.

     

Local thermal sensation and finger vasoconstriction in the locally heated hand

Summary The effects of local heating on finger blood flow (BF) and local thermal sensation (Sens _w ) were studied. Finger BFs in both hands were measured simultaneously; one hand was immersed in water the temperature (T _w ) of which was raised from 35° C to 43°C by steps of 2° C every 10 min, while the other hand was kept atT _w 35°C. FingerBF in the locally heated hand decreased atT _w 37 to 41°C, while fingerBF in the control hand did not alter. Sens_w, in the heated hand showed a dynamic response, initially increasing concomitantly with an increase inT _w , then gradually returning and adapting to a new level of Sens_w. The dynamic response of Sens_w, was not perceived during mental calculation even whenT _w was raised to 40°C, and the reduction in finger blood flow was not observed. These results suggest that finger vasoconstriction caused by local heating closely relates to the dynamic response characteristic of local thermal sensation atT _w above core temperature, and that the perception of local thermal sensation in the central nervous system is involved in the mechanism of this vasoconstrictor response.     

Genetic variability of sexual behavior in a natural population ofDrosophila melanogaster

Although many experiments on laboratory stocks ofDrosophila have suggested that mate choice is a major feature of sexual selection in this organism, few attempts have been made to measure its extent in wild populations. In this study, a crossing design was used to obtain a set of 13 genetically identical independent lines representative of genotypes from an African population ofDrosophila melanogaster. They were tested for variation in sexual behavior using dyadic tests. Significant variation in orientation and vibration latencies was found for males, and in mating speed and copulation duration for both sexes. No evidence of assortative mating, either positive or negative, was found. The absence of a correlation in mating speed between males and females sharing the same genotype leads us to doubt the applicability of the notion of male eagerness and female reluctancy inDrosophila and the importance of vigor as a factor in mating speed. The absence of mate choice in natural populations ofDrosophila seems to us the most likely hypothesis on the basis of both theory and empirical evidence.     

T helper type 2-like cells and therapeutic effects of interferon-γ in combined immunodeficiency with hypereosinophilia (Omenn's syndrome)

We characterized the defects of CD4+ cells in a 17-month-old girl suffering from combined immunodeficiency with hypereosinophilia (Omenn's syndrome). Because the vast majority of peripheral blood CD4⁺ cells expressed the CD45R0 isoform, we purified circulating CD4⁺ CD45R0⁺ cells from the patient and healthy individuals in order to compare their production of cytokines. The patient's CD4⁺ CD45R0⁺ cells spontaneously produced high levels of interleukin-5 (IL-5) in vitro (1600 pg/ml after 24 h of culture) and this was associated with the presence of IL-5 in serum (323 pg/ml). After stimulation with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187, they produced higher levels of IL-4 (306 vs. 55 ± 4 pg/ml) and IL-5 (2900 vs. 213 ± 72 pg/ml) and lower levels of IL-2 (17 vs. 63 ± 17 IU/ml) and interferon-γ (IFN-γ) (16 vs. 299 ± 70 IU/ml) than controls CD4⁺ CD45R0⁺ cells. This T helper type 2 (Th2) pattern was confirmed by the detection using reverse polymerase chain reaction of IL-4, IL-5 and IL-10 mRNA within peripheral blood mononuclear cells. During a therapeutic trial with human IFN-γ (40 μg/day) which ameliorated the clinical status of the patient, we observed a down-regulation of the in vivo expression of IL-5 and IL-10, a normalization of the eosinophil count and an improvement of the Tcell response to phytohemagglutinin. This observation indicates for the first time that Th2-like cells might be involved in certain forms of congenital immunodeficiency and that IFN-γ might down-regulate their activities in vivo.     

Clostridium difficile vaccine and serum immunoglobulin G antibody response to toxin A

There is a strong association between serum antibody responses to toxin A and protection against Clostridium difficile diarrhea. A parenteral C. difficile toxoid vaccine induced very-high-level responses to anti-toxin A immunoglobulin G (IgG) in the sera of healthy volunteers. After vaccination, the concentrations of anti-toxin A IgG in the sera of all 30 recipients exceeded the concentrations that were associated with protection in previous clinical studies. Furthermore, the median concentration of serum anti-toxin A IgG in the test group was 50-fold higher than the previous threshold. These findings support the feasibility of using a vaccine to protect high-risk individuals against C. difficile-associated diarrhea and colitis.     

Impaired responses to toll-like receptor 4 and toll-like receptor 3 ligands in human cord blood

Toll-like receptor (TLR)-4 signaling pathway plays an essential role in host defense against gram-negative bacteria while TLR-3-mediated signaling is critically involved in anti-viral immunity. To gain insight into the defects responsible for impaired Th1 responses in human newborns, we investigated the responses of human cord blood cells to lipopolysaccharide, LPS, and to polyinosinic-polycytidylic acid, Poly (I:C), ligands of TLR-4 and TLR-3, respectively. Measurement of cytokine levels revealed a profound defect in IL-12 (p70) synthesis and an increased release of IL-10 in cord blood exposed to LPS or Poly (I:C), as compared to adult blood. Moreover, Poly (I:C)-induced IFN-alpha production was found to be significantly impaired in cord blood. Phenotypic maturation of myeloid DC in response to LPS or Poly (I:C) was next compared in cord and adult blood. We observed that neonatal myeloid DC displayed decreased upregulation of CD40, CD80 whereas CD86 and HLA-DR upregulation did not differ significantly between adults and neonates. Taken together, these findings might be relevant to the increased vulnerability of human newborns to intracellular pathogens and to their inability to develop efficient Th1-type responses.