Linkage of the MHC to familial multiple sclerosis suggests genetic heterogeneity. The Multiple Sclerosis Genetics Group

Multiple sclerosis (MS) is a demyelinating autoimmune disease of the central nervous system. While its etiology is not well understood, genetic factors are clearly involved. Until recently, most genetic studies in MS have been association studies using the case-control design testing specific candidate genes and studying only sporadic cases. The only consistently replicated finding has been an association with the HLA-DR2 allele within the major histocompatibility complex (MHC) on chromosome 6. Using the genetic linkage design, however, evidence for and against linkage of the MHC to MS has been found, fostering suggestions that sporadic and familial MS have different etiologies. Most recently, two of four genomic screens demonstrated linkage to the MHC, although specific allelic associations were not tested. Here, a dataset of 98 multiplex families was studied to test for an association to the HLA-DR2 allele in familial MS and to determine if genetic linkage to the MHC was due solely to such an association. Three highly polymorphic markers (HLA-DR, D6S273 and TNFbeta) in the MHC demonstrated strong genetic linkage (parametric lod scores of 4.60, 2.20 and 1.24, respectively) and a specific association with the HLA-DR2 allele was confirmed (TDT; P < 0.001). Stratifying the results by HLA-DR2 status showed that the linkage results were limited to families segregating HLA-DR2 alleles. These results demonstrate that genetic linkage to the MHC can be explained by the HLA-DR2 allelic association. They also indicate that sporadic and familial MS share a common genetic susceptibility. In addition, preliminary calculations suggest that the MHC explains between 17 and 62% of the genetic etiology of MS. This heterogeneity is also supported by the minority of families showing no linkage or association with loci within the MHC.      

The phenotype of human placental macrophages and its variation with gestational age

The antigenic phenotype of human villous stromal macrophages (M phi s) from first and third trimester placentas was analyzed using a large number of monoclonal antibodies (MAbs) to monocyte (Mo)/M phi-associated cell membrane determinants. The purpose of this study was to investigate M phi phenotypic heterogeneity to create a database for the correlation of M phi phenotype with specific immunologic functions. The results showed that villous stromal mononuclear cells express many cell surface antigens found on Mo and M phi s and that they are morphologically diverse, ranging in appearance from classic Hofbauer cells to spindle-shaped cells with long cytoplasmic processes. Villous stromal M phi s were the numerically dominant cell type in this structure and exhibited some major phenotypic differences from M phi s in other tissues. Comparison of first- and third-trimester placentas revealed variation in antigen expression with increasing gestational age, in particular of class II major histocompatibility complex (MHC) determinants: HLA-DR and HLA-DP antigen density was low on first-trimester villous M phi s and much higher on third-trimester M phi s while HLA-DQ was undetectable in the first trimester but present on cells in third trimester placentas. The CD1 (T6) antigen, found on Langerhans (LH) cells and cortical thymocytes, was detected on villous M phi s by two thirds of the MAbs directed against different epitopes on this determinant. Furthermore, comparison with similar studies of lymphoid tissues showed that villous M phi s and dendritic cells share the expression of a number of other cell surface antigens. Finally, it was shown that M phi s in first- and third-trimester villi exhibit strong reactivity with MAbs (Leu 3a,b) to the CD4 antigen that serves as the receptor for the human immunodeficiency virus (HIV), suggesting that these cells may be a portal of entry or reservoir for this virus in the fetuses of pregnant, HIV+ women.     

Typing of clinical herpes simplex virus isolates with mouse monoclonal antibodies to herpes simplex virus types 1 and 2: comparison with type-specific rabbit antisera and restriction endonuclease analysis of viral DNA

A total of 122 clinical isolates of herpes simplex virus (HSV) from 107 patients were typed by using an indirect immunoperoxidase technique with commercially available type-specific rabbit antisera, recently developed mouse monoclonal antibodies to HSV types 1 and 2, and restriction endonuclease analysis of viral DNA. With the commercially available type-specific rabbit antisera, 34% of clinical HSV isolates were of indeterminate type; 63% of them were typed as HSV type 1 and 37% as HSV type 2 by using monoclonal antibody and restriction enzyme typing systems. Typing by immunofluorescence assay with the monoclonal antibodies gave identical results to those obtained by restriction enzyme analysis. Simultaneous infection with both HSV types was demonstrated by monoclonal antibody typing in five isolates from three patients. These findings were subsequently confirmed by plaque purification and restriction endonuclease analysis of viral DNA. Monoclonal antibodies were as sensitive as restriction enzyme analysis for the typing of clinical HSV isolates. Because of their simplicity, they are more amenable to use in clinical laboratories than is restriction endonuclease analysis.     

Lipopolysaccharide (LPS) from Brucella abortus is less toxic than that from Escherichia coli, suggesting the possible use of B. abortus or LPS from B. abortus as a carrier in vaccines.

Brucella abortus may be useful as a component of vaccines. This is because it possesses several unique properties as a carrier that enable it to stimulate human B cells even in the relative absence of T cells. Human immunodeficiency virus type 1 proteins conjugated to B. abortus could induce neutralizing antibodies against human immunodeficiency virus type 1. Recently we showed that the characteristics of lipopolysaccharide (LPS) derived from B. abortus are similar to those of the whole bacterium in that the LPS acts as a T-independent type 1 carrier in mice. In this study we wanted to determine whether LPS derived from B. abortus is associated with the adverse effects seen with other bacterial endotoxins. LPS purified from B. abortus by butanol extraction was shown to have less than 2% (wt/wt) contamination by protein and less than 1% (wt/wt) contamination by nucleic acids and to contain 1% (wt/wt) ketodeoxyoctanic acid. Compared with LPS derived from Escherichia coli, B. abortus LPS was 10,000-fold less potent in eliciting fever in rabbits, 268-fold less potent in killing D-galactosamine-sensitized mice, and 1,400-fold and 400-fold less potent in inducing interleukin-1 beta and tumor necrosis factor alpha production, respectively. These results suggest that B. abortus LPS is much less likely than the LPS from E. coli to evoke endotoxic shock; therefore, it may be feasible to incorporate B. abortus as a component of vaccines.     

Comparative activity of teicoplanin and vancomycin against 400 penicillin susceptible and resistantStreptococcus pneumoniae

The MICs of teicoplanin and vancomycin were determined for 400 penicillin-susceptible and -resistant strains ofStreptococcus pneumoniae isolated during a multicenter study in 1992. Teicoplanin displayed a four-fold better activity than vancomycin, with modal MICs of these agents being 0.06 and 0.25 µg/ml, respectively. These data warrant further studies with teicoplanin in the treatment of pneumococcal infections.J. Akli, Centre Hospitalier Général, 41016 Blois Cédex; P. Allouch, Hôpital André Mignot, 78157 Le Chesnay Cédex; C. Bébéar, Centre Hospitalier Pellegrin-Tripode, 33076 Bordeaux Cédex; P. Blondel, Centre Hospitalier Général, 93205 Saint-Denis Cédex 1; A. Boisivon, Centre Hospitalier Général, 78104 Saint-Germain en Laye; J.C. Burdin, Hôpital Central, 54035 Nancy Cédex; H. Chardon, Centre Hospitalier Général, 13616 Aix en Provence Cédex; A. Coupry, Nouvel Hôpital de Fleyrat, 01012 Bourg-en-Bresse Cédex; J.C. Croix, Centre Hospitalier Général, 10003 Troyes Cédex; G. Dorche, Hôpital Bellevue, 42043 Saint-Etienne Cédex; P. Fiévet, Centre Hospitalier, 59507 Douai Cédex; F. Geffroy, Centre Hospitalier René Laënnec, 29107 Quimper Cédex; P. Geslin, Centre Hospitalier Intercommunal, 94010 Créteil; M.L. Grillot, Hôpital Général, 76083 Le Havre Cédex; B. Hautefort, Centre Hospitalier Général, 13697 Arles Cédex; M. Larrouy, Centre Hospitalier Intercommunal de la Côte Basque, 64109 Bayonne; H. Lefrand, Hôpital Henri Duffaut, 84902 Avignon Cédex 9; J.F. Lemeland, Hôtel-Dieu, 76000 Rouen; J.M. Libert, Centre Hospitalier Mane-Lannelongue, 92350 Le Plesssis-Robinson; A. Marmonier, Centre Hospitalier, 72037 Le Mans Cédex; M. Melon, Centre Hospitalier Général, 64011 Pau Cédex; M.H. Nicolas, Hôpital Ambroise Paré, 92104 Boulogne; J.C. Reveil, Hôpital Manchester, 08011 Charleville-Mézières Cédex; Y. Rio, Hôpital Notre Dame de Bon Secours, 57038 Metz Cédex; R. Sanchez, Hôpital Général, 24019 Périgueux Cédex; A. Sédaillan, Centre Hospitalier, 74011 Annecy Cédex, France.     

Relationship of resting blood pressure and heart rate to experienced anger and expressed anger

Forty-five nonmedicated subjects rated on analog scales the anger they experienced at home and at work (Experienced Anger). They also rated the extent to which others were aware of their anger (Expressed Anger) and the extent to which anger had been expressed in their families or origin (Family Expressed Anger). They were then physiologically monitored during a 2-min relaxation period that followed habituation to the laboratory setting. For the group as a whole, Expressed Anger was inversely related to systolic (SBP) and diastolic (DBP) blood pressure while Family Expressed Anger was inversely related to SBP only. When the sample was divided into normotensive and hypertensive subgroups, the normotensives showed significant associations between Experienced Anger and SBP, Expressed Anger and DBP, and Family Expressed Anger and SBP. The hypertensive subgroup showed no significant associations. Both males and females showed a significant association between Expressed Anger and DBP, but only females showed this relationship with SBP. It is concluded that coping with anger by conscious inhibition of its expression is associated with increases in both systolic and diastolic blood pressure.     

The BPV-1 E5 oncoprotein expressed in Schizosaccharomyces pombe exhibits normal biochemical properties and binds to the endogenous 16-kDa component of the vacuolar proton-ATPase

The 44-amino-acid E5 oncoprotein of bovine papillomavirus type 1 transforms immortalized murine fibroblast cell lines. This highly hydrophobic protein forms homodimers, localizes to intracellular membrane compartments (including the Golgi apparatus), and forms a complex with the 16-kDa membrane-embedded constituent (16k) of the vacuolar proton-ATPase. To develop a system for the genetic and biochemical analysis of the E5/16k interaction, the E5 gene was cloned into a new vector which was designed for expression in the fission yeast Schizosaccharomyces pombe. The E5 protein synthesized in this system dimerized normally and bound to endogenous and overexpressed S. pombe 16k protein. Comparison of the S. pombe and mammalian 16k proteins showed strong conservation in carboxyl-terminal amino acids but greater variation in the amino-terminal sequences, suggesting that E5 was interacting with the 16k carboxyl domains. Finally, a new protein epitope tag is described which permitted for the first time the coprecipitation of E5 with antibodies directed against the 16k protein.     

Identifying candidate causal variants responsible for altered activity of the ABCB1 multidrug resistance gene

The difficulty of fine localizing the polymorphisms responsible for genotype-phenotype correlations is emerging as an important constraint in the implementation and interpretation of genetic association studies, and calls for the definition of protocols for the follow-up of associated variants. One recent example is the 3435C>T polymorphism in the multidrug transporter gene ABCB1, associated with protein expression and activity, and with several clinical conditions. Available data suggest that 3435C>T may not directly cause altered transport activity, but may be associated with one or more causal variants in the poorly characterized stretch of linkage disequilibrium (LD) surrounding it. Here we describe a strategy for the follow-up of reported associations, including a Bayesian formalization of the associated interval concept previously described by Goldstein. We focus on the region of high LD around 3435C>T to compile an exhaustive list of variants by (1) using a relatively coarse set of marker typings to assess the pattern of LD, and (2) resequencing derived and ancestral chromosomes at 3435C>T through the associated interval. We identified three intronic sites that are strongly associated with the 3435C>T polymorphism. One of them is associated with multidrug resistance in patients with epilepsy (chi2 = 3.78, P = 0.052), and sits within a stretch of significant evolutionary conservation. We argue that these variants represent additional candidates for influencing multidrug resistance due to P-glycoprotein activity, with the IVS 26+80 T>C being the best candidate among the three intronic sites. Finally, we describe a set of six haplotype tagging single-nucleotide polymorphisms that represent common ABCB1 variation surrounding 3435C>T in Europeans.