The BALB/c mouse B-cell response to pigeon cytochrome c initiates as a heteroclitic response specific for the self antigen mouse cytochrome c.

Direct evidence is presented in support of the longstanding but unproven hypothesis that B lymphocytes specific for self antigens (Ags) can be used in the immune response to foreign Ags. We show that the B cells in BALB/c mic responding early to pigeon cytochrome c (CYT) produce antibodies that recognize and bind the major antigenic site on mouse CYT with greater affinity than they bind pigeon CYT i.e., they are heteroclitic for the self Ag. Furthermore, these B cells express the same combination of immunoglobulin variable region (V) genes that are known to be used in B-cell recognition of mouse CYT. Over time, the response to pigeon CYT becomes more specific for the foreign Ag through the recruitment of B cells expressing different combinations of V genes and, possibly, somatic mutation of the mouse CYT specific B cells from early in the response. Cross-recognition of pigeon CYT by mouse CYT-specific B cells results from the sharing of critical amino acid residues by the two Ags. Although B-cell recognition of the self Ag, mouse CYT, is very specific, which limits the extent to which foreign Ags can cross-activate the autoreactive B cells, it is possible that polyreactive B cells to other self Ags may be used more frequently in response to foreign Ags.

FAS is highly expressed in the germinal center but is not required for regulation of the B-cell response to antigen.

In establishing the memory B-cell population and maintaining self-tolerance during an immune response, apoptosis mediates the removal of early, low-affinity antibody-forming cells, unselected germinal center (GC) cells, and, potentially, self-reactive B cells. To address the role of the apoptosis-signaling cell surface molecule FAS in the B-cell response to antigen, we have examined the T-cell-dependent B-cell response to the carrier-conjugated hapten (4-hydroxy-3-nitrophenyl)acetyl (NP) in lpr mice in which the fas gene is mutated. High levels of FAS were expressed on normal GC B cells but the absence of FAS did not perturb the progressive decline in numbers of either GC B cells or extrafollicular antibody-forming cells. Furthermore, the rate of formation and eventual size of the NP-specific memory B-cell population in lpr mice were normal. The accumulation of cells with affinity-enhancing mutations and the appearance of high-affinity anti-NP IgG1 antibody in the serum were also normal in lpr mice. Thus, although high levels of FAS are expressed on GC B cells, FAS is not required for GC selection or for regulation of the major antigen-specific B-cell compartments. The results suggest that the size and composition of B-cell compartments in the humoral immune response are regulated by mechanisms that do not require FAS.

Fine specificity of the antibody response to myelin basic protein in the central nervous system in multiple sclerosis: the minimal B-cell epitope and a model of its features.

T cells, B cells, and antibody are found in the white matter of the central nervous system in multiple sclerosis. The epitope center for the antibody response to human myelin basic protein (MBP) fits precisely the minimal epitope Pro85-Val-Val-His-Phe-Phe-Lys-Asn-Ile-Val-Thr-Pro96 for that reported for HLA DR2b (DRB1*1501)-restricted T cells that recognize MBP [Wucherpfenning, K.W., Sette, A., Southwood, S., Oseroff, C., Matsui, M., Strominger, J. & Hafler, D. (1994) J. Exp. Med. 179, 279-290], and overlaps with the reported DR2a-restricted epitope for T cells reactive to MBP [Martin, R., Howell, M. D., Jaraquemada, D., Furlage, M., Richert, J., Brostoff, S., Long, E. O., McFarlin, D. E. & McFarland, H. F. (1991) J. Exp. Med. 173, 19-24]. We describe a molecular model of this epitope.

X-ray structure of clotting factor IXa: active site and module structure related to Xase activity and hemophilia B.

Hereditary deficiency of factor IXa (fIXa), a key enzyme in blood coagulation, causes hemophilia B, a severe X chromosome-linked bleeding disorder afflicting 1 in 30,000 males; clinical studies have identified nearly 500 deleterious variants. The x-ray structure of porcine fIXa described here shows the atomic origins of the disease, while the spatial distribution of mutation sites suggests a structural model for factor X activation by phospholipid-bound fIXa and cofactor VIIIa. The 3.0-A-resolution diffraction data clearly show the structures of the serine proteinase module and the two preceding epidermal growth factor (EGF)-like modules; the N-terminal Gla module is partially disordered. The catalytic module, with covalent inhibitor D-Phe-1I-Pro-2I-Arg-3I chloromethyl ketone, most closely resembles fXa but differs significantly at several positions. Particularly noteworthy is the strained conformation of Glu-388, a residue strictly conserved in known fIXa sequences but conserved as Gly among other trypsin-like serine proteinases. Flexibility apparent in electron density together with modeling studies suggests that this may cause incomplete active site formation, even after zymogen, and hence the low catalytic activity of fIXa. The principal axes of the oblong EGF-like domains define an angle of 110 degrees, stabilized by a strictly conserved and fIX-specific interdomain salt bridge. The disorder of the Gla module, whose hydrophobic helix is apparent in electron density, can be attributed to the absence of calcium in the crystals; we have modeled the Gla module in its calcium form by using prothrombin fragment 1. The arched module arrangement agrees with fluorescence energy transfer experiments. Most hemophilic mutation sites of surface fIX residues occur on the concave surface of the bent molecule and suggest a plausible model for the membrane-bound ternary fIXa-FVIIIa-fX complex structure: fIXa and an equivalently arranged fX arch across an underlying fVIIIa subdomain from opposite sides; the stabilizing fVIIIa interactions force the catalytic modules together, completing fIXa active site formation and catalytic enhancement.

DNA-mediated immunization to the hepatitis B surface antigen in mice: aspects of the humoral response mimic hepatitis B viral infection in humans.

Intramuscular injection of plasmid DNA expression vectors encoding the three envelope proteins of the hepatitis B virus (HBV) induced humoral responses in C57BL/6 mice specific to several antigenic determinants of the viral envelope. The first antibodies appeared within 1-2 weeks after injection of DNA and included antibodies of the IgM isotype. Over the next few weeks, an IgM to IgG class switch occurred, indicating helper T-lymphocyte activity. Peak IgG titers were reached by 4-8 weeks after a single DNA injection and were maintained for at least 6 months without further DNA injections. The antibodies to the envelope proteins reacted with group- and subtype-specific antigenic determinants of the HBV surface antigen (HBsAg). Expression vectors encoding the major (S) and middle (preS2 plus S) envelope proteins induced antibodies specific to the S protein and preS2 domain, and preS2 antibodies were prominent at early time points. In general, the expression vectors induced humoral responses in mice that mimic those observed in humans during the course of natural HBV infection.

Blockade of T- and B-lymphocyte development by antibody to the gamma c subunit of the receptors for interleukins 2, 4, and 7.

Cytokines are important regulators of hematopoesis. Mutations in gamma c, which is a subunit shared by the receptors for interleukin (IL) 2, IL-4, and IL-7, have been causally associated with human X chromosome-linked severe combined immunodeficiency disease. This finding indicates a mandatory role for cytokine receptor signaling at one or more stages of lymphocyte development. To evaluate the cellular level at which gamma c is critical for lymphopoiesis, the effect of monoclonal antibodies to gamma c on the capacity of syngeneic bone marrow cells to reconstitute the hematopoietic compartment of lethally irradiated recipient mice was examined. We show that monoclonal antibody to gamma c blocked lymphocyte development at or before the appearance of pro-B cells and prior to or at the seeding of the thymus by precursor cells while erythromyeloid cell development was normal. These results suggest that one level of lymphocyte development that requires gamma c is a point in hematopoietic cell differentiation near the divergence of lymphopoiesis and erythromyelopoesis.

A cyclophilin from the polycentric anaerobic rumen fungus Orpinomyces sp. strain PC-2 is highly homologous to vertebrate cyclophilin B.

A cyclophilin (CyP) purified to homogeneity from the polycentric anaerobic rumen fungus Orpinomyces sp. strain PC-2 had a molecular mass of 20.5 kDa and a pI of 8.1. The protein catalyzed the isomerization of the prolyl peptide bond of N-succinyl-Ala-Ala-(cis,trans)-Pro-Phe p-nitroanilide with a kcat/Km value of 9.3 x 10(6) M-1.s-1 at 10 degrees C and pH 7.8. Cyclosporin A strongly inhibited this peptidylprolyl cis-trans isomerase activity with an IC50 of 19.6 nM. The sequence of the first 30 N-terminal amino acids of this CyP had high homology with the N-terminal sequences of other eukaryotic CyPs. By use of a DNA hybridization probe amplified by PCR with degenerate oligonucleotide primers designed based on the amino acid sequences of the N terminus of this CyP and highly conserved internal regions of other CyPs, a full-length cDNA clone was isolated. It possessed an open reading frame encoding a polypeptide of 203 amino acids with a calculated molecular weight of 21,969, containing a putative hydrophobic signal peptide sequence of 22 amino acids preceding the N terminus of the mature enzyme and a C-terminal sequence, Lys-Ala-Glu-Leu, characteristic of an endoplasmic reticulum retention signal. The Orpinomyces PC-2 CyP is a typical type B CyP. The amino acid sequence of the Orpinomyces CyP exhibits striking degrees of identity with the corresponding human (70%), bovine (69%), mouse (68%), chicken (66%), maize (61%), and yeast (54%) proteins. Phylogenetic analysis based on the CyP sequences indicated that the evolutionary origin of the Orpinomyces CyP was closely related with CyPs of animals.

La narrativa en La Ilustración (1849-1857): la «Serie B» del Semanario Pintoresco Español

En los ocho años de vida de La Ilustración, la narrativa, larga y breve, fue uno de los elementos fundamentales de este periódico ilustrado. El artículo analiza los relatos que allí aparecieron y los efectos que las necesidades industriales y comerciales tuvieron en una revista que luchaba por sobrevivir económicamente. Ángel Fernández de los Ríos, propietario del Semanario Pintoresco Español y de La Ilustración, convirtió a esta última, en lo que toca a la narrativa, en una especie de serie B, en la que se entremezclaban reediciones, traducciones y colaboraciones que no habían llegado a tener el nivel suficiente para llegar a las páginas del Semanario Pintoresco Español. La abundancia de reediciones y las diversas fuentes de las que proceden hacen ver que en su selección fue más importante la imperiosa necesidad de sacar la publicación a la calle cada quince días que otros criterios. Ello también explica la abundancia de traducciones de novelas y cuentos en las páginas de La Ilustración. Traducciones y reediciones son el cuerpo principal de la narrativa publicada en La Ilustración y si tenemos que detenernos en las obras de más calidad que allí aparecen, será en este tipo de relatos, puesto que los textos narrativos originales, muy a menudo publicados sin firma de autor, resultan de muy escaso interés. Solo podemos contra con la excepción de dos relatos de José Güell y Renté (Anacaona y Quibiam) y uno de Gertrudis Gómez de Avellaneda: La baronesa de Joux. No representa, pues, La Ilustración una fuente de importancia para la narración española del medio siglo. Su posición subordinada al Semanario Pintoresco Español impidió que la publicación desarrollara una línea de narraciones de calidad. Es testimonio, sin embargo, de un proceso de industrialización de la labor periodística y literaria que estaba desarrollándose con gran celeridad si tenemos en cuenta el fracaso de El Artista y del No me olvides apenas quince años antes. A la altura del medio siglo el periodismo literario se había convertido en una industria que exigía ingentes esfuerzos de trabajo y una producción torrencial.

Letter from William Croswell to Mr. B, [ca. 1829]

Draft of a letter regarding Croswell's study of the New Testament.

B. Copies of three letters to Moody Noyes, 1800

It is unknown who made these manuscript copies of three letters from John Henry Tudor to Moody Noyes; they are not in Tudor's hand. The letters were written on September 23, 1800; November 7, 1800; and February 20, 1801. Noyes and Tudor were classmates at Harvard College, where both graduated in the class of 1800. The letters were written after they had graduated from Harvard, and in them Tudor recounts travels with his family around New England, including a stay in Saratoga and Ballston Springs, New York; his interest, shared by Moody, in entering into a store or other form of business, although he found "merchants in general [to be] a contemptible set of beings"; the maxims of the Duke de la Rochefoucauld; his hurt feelings at Moody's failure to answer his letters; and his imminent travels to Cuba with his brother, Frederic, made in hopes of restoring his health.

Studies of the waters of the continental shelf, Cape Cod to Chesapeake Bay : III. A volumetric study of the zooplankton / by Henry B. Bigelow and Mary Sears.

v.54:no.4 (1939)

L.B.'s letter respecting excuse to president of College, 1797 August 31

Brief handwritten note to Loammi Baldwin from his son requesting a written excuse to be sent to the Harvard President for his absence on election day.

B. Copies of three letters to Moody Noyes, 1800

It is unknown who made these manuscript copies of three letters from John Henry Tudor to Moody Noyes; they are not in Tudor's hand. The letters were written on September 23, 1800; November 7, 1800; and February 20, 1801. Noyes and Tudor were classmates at Harvard College, where both graduated in the class of 1800. The letters were written after they had graduated from Harvard, and in them Tudor recounts travels with his family around New England, including a stay in Saratoga and Ballston Springs, New York; his interest, shared by Moody, in entering into a store or other form of business, although he found "merchants in general [to be] a contemptible set of beings"; the maxims of the Duke de la Rochefoucauld; his hurt feelings at Moody's failure to answer his letters; and his imminent travels to Cuba with his brother, Frederic, made in hopes of restoring his health.