Virion-bound ICAM-1 and activated LFA-1: a combination of factors conferring resistance to neutralization by sera from human immunodeficiency virus type 1-infected individuals independently of the disease status and phase

The role of the supplementary interaction between virion-bound host ICAM-1 and LFA-1 on target cells in sensitivity to neutralization of human immunodeficiency virus type 1 (HIV-1) is poorly studied. Serum samples from four long-term nonprogressors (LTNPs) and sequential sera from one progressor were used to assess neutralization sensitivity of isogenic ICAM-1-negative and ICAM-1-bearing HIV-1(NL4-3), a prototype of T-cell-line-adapted viruses. We found that virus neutralization sensitivity to the studied sera is not modified by the additional interaction between virally embedded ICAM-1 and LFA-1 under an inactive state. However, expression on the target cell surface of an activated LFA-1 form renders ICAM-1-bearing virus particles, but not viruses devoid of ICAM-1, more refractory to neutralization by sera from three out of four LTNPs and all sequential sera from the person who has experienced a progression of the HIV-1-associated disease. Although no conclusive correlation could be drawn between virus susceptibility to neutralization and the disease status or stages of HIV-1 infection, these findings demonstrate that other nonspecific virus-cell interactions mediated by virion-anchored host proteins and their normal cognate ligands on target cells represent factors that can affect the mechanism of HIV-1 neutralization.     

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.     

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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Electronic Supplementary Material The french version of this article is available in the form of electronic supplementary material and can be obtained by using the Springer Link server located at

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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.     

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.     

Sodium gradient-energized concentrative transport of adenosine in renal brush border vesicles

The uptake of adenosine in brush border vesicles of the proximal tubule of the rat kidney has been studied with a filtration technique. The initial rate of uptake was almost 6 times greater in the presence of NaCl than in the presence of KCl. The stimulatory effect of Na⁺ was strictly dependent on a gradient of Na⁺ (out>in). The time course of uptake showed an overshoot with a maximum at 20 s with a gradient of NaCl, but not with KCl. Inosine and 5-AMP were produced from adenosine within the vesicles. In the presence of an inhibitor or adenosine deaminase adenosine was not significantly metabolized during the first 20 s of uptake. Thus, kinetic parameters of transport could be studied in the absence of interferences with metabolism. AK _m of 1.1 M and aV _max of 232 pmol · min^–1 · mg protein^–1 were calculated for the Na⁺ gradient-dependent transport. The dependency on a Na⁺ gradient, the capacity for uphill transport and the high affinity for adenosine situate this transport system apart from the mechanisms of transport of nucleosides described so far. It may be relevant in regard to the role of adenosine in the regulation of glomerular filtration.Abbreviations used EHNA erythro-9-(2-hydroxy-3-nonyl)adenine - FCCP carbonylcyanide p-trifluoromethoxy-phenylhydrazone - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - Tris tris (hydroxymethyl)-aminomethane     

Cloned alpha and beta C-protein antigens of group B streptococci elicit protective immunity.

Streptococcus agalactiae (group B streptococci [GBS]) is the leading cause of neonatal sepsis and meningitis in the United States. The surface-associated C proteins of GBS play a role in immunity, but their number, size, structure, function, and virulence properties have not been well characterized. A recombinant library of DNA fragments from GBS strain A909 (type Ia/C) was prepared in the plasmid pUX12, a specially constructed Escherichia coli expression vector. The library was screened with a rabbit antiserum shown to be protective for passive immunity to GBS infection in a mouse lethality model. Clones were divided into two distinct groups on the basis of DNA-DNA cross-hybridization, restriction enzyme analysis, and the expression of antigenic proteins in E. coli. A characteristic clone from each group was chosen for further study. Clone pJMS23 expresses gene products that biochemically and immunologically correspond to the trypsin-resistant, C-protein alpha antigen. Clone pJMS1 expresses a gene product that binds to immunoglobulin A and is similar to the trypsin-sensitive, C-protein beta antigen. Antisera raised in rabbits against E. coli containing each of the plasmid clones were able to elicit protective immunity in mice challenged by GBS strains carrying the C proteins but not by non-C-protein-bearing strains. Southern blot analysis shows no DNA homology between the clones, and there is no immunological cross-reactivity between the antigens they express. Therefore, pJMS23 and pJMS1 encode two different C proteins that define unique protective epitopes.