Showing papers by "New York Blood Center published in 1988"

Failure of a human immunodeficiency virus (HIV) immune globulin to protect chimpanzees against experimental challenge with HIV.

Abstract: To assess the possible efficacy of passive immunization against human immunodeficiency virus (HIV) an immune globulin was prepared from plasma of HIV-seropositive donors selected to be among those having the top 12.5% of virus-neutralizing antibody titers. The immune globulin was treated with pepsin to render it intravenously tolerable. The preparation, which we termed HIVIG, neutralized 100 tissue culture 50% infective doses (TCID50) of HIV at an average dilution of 1:1000 in neutralization tests in vitro. During preparation HIVIG was subjected to virus inactivation and removal procedures that in theory resulted in a reduction in HIV infectivity by a factor of 10(25). At a dose of 9-10 ml/kg of body weight both the virus-inactivated source plasma and the final immunoglobulin preparation were noninfective and without adverse effect in two chimpanzees. Two chimpanzees inoculated intravenously with HIVIG at 1 ml/kg and two inoculated with 10 ml/kg were challenged intravenously 1 day later with 400 TCID50 of the same strain of HIV (HTLV-IIIb) used in neutralization assays in vitro. All animals became infected. Incubation periods to virus isolation (by cocultivation with human mononuclear cells) in HIVIG recipients did not differ significantly from the incubation period seen in a control animal that received a normal anti-HIV-free immunoglobulin. These findings may have implications for understanding the failure of experimental vaccines to protect against HIV challenge in chimpanzee experiments.

A physical map of the human regulator of complement activation gene cluster linking the complement genes CR1, CR2, DAF, and C4BP.

Abstract: We report the organization of the human genes encoding the complement components C4-binding protein (C4BP), C3b/C4b receptor (CR1), decay accelerating factor (DAF), and C3dg receptor (CR2) within the regulator of complement activation (RCA) gene cluster. Using pulsed field gel electrophoresis analysis these genes have been physically linked and aligned as CR1-CR2-DAF-C4BP in an 800-kb DNA segment. The very tight linkage between the CR1 and the C4BP loci, contrasted with the relative long DNA distance between these genes, suggests the existence of mechanisms interfering with recombination within the RCA gene cluster.

The Pre-S Region of Hepadnavirus Envelope Proteins

Abstract: Publisher Summary The biosynthesis and biological properties of hepadnavirus envelope (env) proteins containing the pre-S region are described in this chapter Abundant experimental evidence indicates that the pre-S region of hepadnavirus env proteins has essential biological functions These are (1) involvement in virus attachment to hepatocytes, (2) elicitation of virus-neutralizing antibodies, (3) high antigenicity and immunogenicity, (4) regulation of the genetic restriction of the immune response, and (5) regulation of the process of virus particle assembly It is also suggested that the biosynthesis of hepatitis B virus (HBV) env proteins containing the pre-S region might differ at different stages of hepatitis B infections In contrast to HBsAg subviral particles, P39/GP42 and GP33/GP36 are substantial and not minor components of the HBV envelope The pre-S region of hepadnavirus env proteins can be mimicked very successfully with synthetic peptide analogs Consequently, such synthetic peptides have become important resources for the development of synthetic vaccines against hepatitis B and for the preparation of diagnostic reagents

Correlation between the anticellular and DNA fragmenting activities of tumor necrosis factor.

Abstract: The treatment of cells sensitive to the anticellular effect of tumor necrosis factor (TNF) with TNF results in a degradation of their cellular DNA into DNA fragments that are multiples of 200 base pairs. TNF treatment of cells resistant to the anticellular effect of TNF, but bearing receptors for TNF, fails to result in any DNA fragmentation. Incubation conditions, such as temperature, the presence of metabolic inhibitors or amino acid deprivation, that modulate the effectiveness of TNF or affect the rate at which TNF exerts its anticellular effect have a similar effect on the ability of the TNF to generate DNA fragments. Thus the TNF-mediated DNA fragmentation and the rate at which it occurs correlates with the rate at which cells respond to the anticellular effect of TNF and, as such, might serve as a marker for the responsiveness of cells to TNF.

Biochemical studies on McLeod phenotype red cells and isolation of Kx antigen.

Abstract: Summary Red cells of the McLeod blood group phenotype have weak Kell antigens, lack Kx antigen and have acanthocytic morphology. We have immunoprecipitated Kell antigens from McLeod red cells and show that they are markers on the same 93 kDa membrane protein that carries Kell antigens on normal red cells. However, as determined by Western immunoblotting, McLeod red cells have a marked deficiency of this protein. We have also studied the near-neighbour relationship of McLeod and common Kell red-cell membrane proteins by cross-linking intrinsic sulphydryl groups by oxidation, catalysed with orthophenanthroline and copper, or by cross-linking amino groups with dimethyl 3,3-dithiobispropionimidate. Results were analysed by diagonal mapping in two-dimensional gels. No abnormalities of membrane protein inter-relationship were detected in McLeod red cells. We have isolated Kx antigen from K0 red cells by immuno-precipitation with human alloimmune anti-Kx serum, isolation of immune complexes from detergent-solubilized cell membranes with protein A-Sepharose and analysis of the eluted immune complex by SDS-PAGE under reducing conditions. Kx antigen is a marker on a red-cell membrane protein of approximately 37 kDa. K0 (Knull) red cells have about twice the amount of Kx antigen as do red cells of common Kell type. McLeod red cells have no detectable Kx antigen by serological tests or by immunoprecipitation.

Inactivation of lipid-enveloped viruses in labile blood derivatives by unsaturated fatty acids.

Abstract: Virus sterilization of blood plasma derivatives by addition of several naturally occurring fatty acids was evaluated using vesicular stomatitis virus and Sindbis virus as markers for lipid-enveloped virus inactivation and human immunodeficiency virus (HIV) Inactivation of greater than or equal to 10(4) tissue culture infectious doses (TCID50) of marker viruses added to antihemophilic factor (AHF) concentrates, with 60-100% retention of AHF activity, was achieved with oleic, 11-eicosenoic, linoleic, linolenic, palmitoleic and arachidonic acids Elaidic, gamma-linolenic, palmitic, and arachidic acids and another fat-soluble compound previously reported to inactivate virus, butylated hydroxytoluene, were less effective A long chain mono- but not a di- or triglyceride also displayed virucidal properties Evaluation of the inactivation of HIV added to an immune globulin solution on exposure to 0033% sodium oleate for 20 min indicated inactivation of greater than or equal to 10(34) TCID50 The degree of virus inactivation depended on the sample composition A favorable balance was achieved between degree of virus inactivation and retention of protein function for AHF concentrate, prothrombin complex concentrate, antithrombin III concentrate, and immune globulin solution on incubation with 0033% (w/v) sodium oleate at 24 degrees C for 4-6 h Virus inactivation in whole plasma and plasma cryoprecipitate was not complete despite use of higher concentrations of sodium oleate and/or incubation at 37 degrees C Reduced virus kill in these less purified derivatives probably is a consequence of their endogenous lipid and/or albumin

Tumor necrosis factor and IFN induce a common set of proteins.

Abstract: The treatment of cells with TNF or IFN results in the development of an antiviral state and in the induction of a common set of proteins with m.w. of 80,000, 67,000, and 56,000. The induction of the 80,000- and 56,000-Da proteins after TNF treatment is dependent on the synthesis of an intermediary protein, whereas the induction of the 67,000-Da protein appears to occur as a direct result of the TNF treatment. The effects of antibodies to IFN on the TNF-mediated effects have been evaluated and reveal that the incubation of TNF-treated cells with antibody to rIFN-beta 1 greatly reduces the antiviral effectiveness of the TNF treatment and blocks the ability of TNF to induce the 80,000-Da protein. Incubation with antibodies to either IFN-alpha or IFN-gamma failed to affect the TNF-mediated responses. Thus, the induction by TNF of each of the proteins is regulated differently and is mediated through both IFN-dependent and IFN-independent mechanisms.

The development of antibody to the interferon-induced indoleamine 2,3-dioxygenase and the study of the regulation of its synthesis.

Abstract: Interferon (IFN) treatment of cells induces the synthesis of several new proteins. Antibody to the IFN-induced 42,000 dalton protein has been prepared and used in the study of this protein. The synthesis of the 42,000 dalton protein is dependent on de novo RNA synthesis, as its induction can be blocked if actinomycin D and the IFN are added to the cells simultaneously. Several lines of evidence suggest that the IFN-induced 42,000 dalton protein is the IFN-induced indoleamine 2,3-dioxygenase (IDO). These are as follows: 1) antibody to the 42,000 dalton protein neutralizes the activity of the IDO; 2) examination of a variety of cell lines reveals a correlation between the presence of this protein and the presence of the IDO; and 3) the induction of both the IDO and the 42,000 dalton protein is blocked under conditions in which the IFN treatment is performed in the presence of cycloheximide, and actinomycin D is added to the cells prior to the removal of the cycloheximide. A study of a variety of cell lines ...

Coagulation factor XIII B subunit is encoded by a gene linked to the regulator of complement activation (RCA) gene cluster in man

Abstract: We have performed linkage analysis to determine the genetic relationship between the loci coding for coagulation factor XIII B (F13B) and the regulator of complement activation (RCA) gene cluster, comprised of the loci encoding C3b/C4b-receptor (CR1), C3d/Epstein-Barr virus-receptor (CR2), decay accelerating factor (DAF), C4b-binding protein (C4BP), and factor H (HF), as the products of these six loci show structural similarities. Here we report that the human F13B gene is also a member of RCA gene cluster and that it maps in close proximity to the gene encoding complement factor H. Coagulation factor XIII is a tetrameric complex composed of two A and two B polypeptides held together by noncovalent bonds (Schwartz et al. 1973, Chung et al. 1974). Factor XIII is activated by the serine protease thrombin which cleaves a 36 amino acid (aa) peptide from the amino terminus of the catalytic A subunit. In the presence of Ca 2+, the activated A' subunit is released from the B subunit and acquires the transglutaminase enzymatic activity which is responsible for the cross-linking of the fibrin monomers (reviewed in Folk and Finlayson 1977, Lorand et al. 1980). The precise function of the B subunit is not completely understood, but it is thought to protect or stabilize the A subunit, regulating the activation of the zymogen (Cooke 1974). The F13B locus displays a genetic polymorphism (Board 1980), and several phenotypic variants have been characterized by isoelectric focusing (IEF) (Dykes and Polesky 1987). Factor XIII B is structurally related to the members of the so-called superfamily of C3b/C4b binding proteins. These proteins have a structural organization based on the presence of internal repeat units of 60 aa which share a framework of highly conserved residues (reviewed in Reid et al. 1986). Factor XIII B is composed of ten of these repeats which represent the amino terminal 98 % of

Children at high risk of diabetes mellitus: New York studies of families with diabetes and of children with congenital rubella syndrome.

Abstract: The frequency of Type I, or insulin-dependent mellitus (IDDM) in the general population is approximately 1 in 600 individuals. By contrast, in families where one child has IDDM, the likelihood that another child will develop IDDM is about 100 times higher. This population, at high risk for the development of IDDM, seemed an ideal group in which to identify some of the factors which might precede the development of clinically apparent IDDM. IDDM is clearly a multifactorial disease. The existence of a gene or genes predisposing to diabetes and mapping within or very near the HLA locus on chromosome six has been suggested by several different investigators. In addition, environmental agents also appear to be needed to “trigger” the predisposition towards overt disease since concordance for monozygotic twins is estimated at only approximately 30%. Convincing evidence has accumulated over the last several years that the mechanism of the expression of the genetic susceptibility is an autoimmune destruction of the pancreatic beta cells.

Delineation of contiguous determinants essential for biological functions of the pre-S sequence of the hepatitis B virus envelope protein: its antigenicity, immunogenicity and cell-receptor recognition.

Abstract: Summary The preS region of the hepatitis B virus (HBV) envelope protein has the following properties: (1) exposure on the surface of the virus; (2) high immunogenicity; (3) involvement in the reaction of the virus with cell receptors and (4) elicitation of antibodies protective against infection. Attempts to mimic B- and T-cell epitopes on the native protein by synthetic peptides were highly successful. This success depended on identification of those regions within the preS sequences which are the most important for biological function of the virus and for immunity, and on the synthesis of long peptides (20–40 residues) containing both B- and T-cell epitopes. Results presented here highlight those subregions of the preS sequence which are the most essential for the antigenicity and immunogenicity of HBV.

Inactivation of viruses in cell- and protein-containing compositions using aryl diol epoxides

Abstract: A process of inactivating infectious viruses in a cell-containing or a protein-containing composition containing such viruses, comprising contacting such composition with an effective amount of at least one aryl diol epoxide of the formula ##STR1## in which X is an aromatic ring system having from 3 to 6 used rings, for a sufficient period of time substantially to inactivate said viruses without incurring substantial disruption or inactivation of cells or without incurring substantial protein denaturation.

Gonadal dimorphism explained as a dosage effect of a locus on the sex chromosomes, the gonad-differentiation locus (GDL).

Abstract: In human somatic cells bearing two X chromosomes, one X is genetically inactivated throughout most of its length, whereas in cells with one X and one Y both sex chromosomes are active (with the exception of the constitutive heterochromatin of the Y that is inert). The vast base of information concerning normal and abnormal human sexual development that has accumulated since the advent of human cytogenetics 3 decades ago can be integrated by the following hypothesis: Homologous gonad-differentiation loci (GDLs) exist on the X and Y. The GDLs are strictly sex-linked; that is, normally they do not recombine during spermatogenesis, so that considerable divergence in DNA sequence doubtless has occurred between the locus on the X and the locus on the Y. The abundance of their evolutionarily conserved product--a substance still to be identified--determines the path of differentiation that the indifferent gonadal anlage of the early embryo will take: if only one GDL is transcribed, the case when two X chromosomes are present, ovary will develop; if two GDLs are transcribed, the case when a Y is present along with an X, testis will develop. By implication, facultative X inactivation is an integral and essential component of the system adopted in mammalian evolution for accomplishing gonadal--viz., sexual--dimorphism.

Gamma-glutamyltransferase as a potential surrogate marker for detection of the non-A, non-B carrier state.

Abstract: Alanine aminotransferase (ALT) is currently widely used as a surrogate marker for detection of blood having a higher risk of transmission of non-A, non-B hepatitis (NANBH). Studies in the chimpanzee model, in which the NANBH carrier state is well defined, have revealed gamma-glutamyltransferase to be considerably more sensitive for the detection of NANBH carrier blood. Further study is required to determine whether this marker is also more sensitive for detection of the chronic NANBH carrier state in man.

Production of a monoclonal antibody directed against an interferon-induced 56,000-dalton protein and its use in the study of this protein.

Abstract: Interferon (IFN) treatment of cells induces the synthesis of several new proteins. A hybridoma cell line producing monoclonal antibody to the IFN-induced 56,000-dalton protein has been developed. The IFN-induced 56,000-dalton protein is synthesized by a variety of different cells and in response to IFN-alpha, IFN-beta, and IFN-gamma. The induction of this protein is dependent on de novo RNA synthesis, since its induction is inhibited if actinomycin D and the IFNs are added to the cells simultaneously. Labeling of IFN-treated cells at 4-h intervals at various times after the addition of the IFNs reveals that the synthesis of the 56,000-dalton protein in IFN-alpha-treated cells peaks within 12 h after the addition of the IFN and is no longer enhanced 20 h after exposure to the IFN. In contrast, IFN-gamma-treated cells continue to show an enhanced synthesis of this IFN-induced protein even after 20 h of exposure to the IFN. Thus, the synthesis of the IFN-induced 56,000-dalton protein is regulated differently by the different IFNs. When cells are treated with IFN-alpha or IFN-gamma in the presence of cycloheximide, and actinomycin D is added prior to the removal of the cycloheximide, the cells produce the IFN-induced 56,000-dalton protein and develop an antiviral state in response to both IFN-alpha and IFN-gamma. These results demonstrate that the synthesis of the 56,000-dalton protein is not dependent on the synthesis of an intermediary protein and that the establishment of an antiviral state occurs in the absence of multiple transcriptional events.

Chimpanzees and AIDS research.

Abstract: Pressure is mounting to relax the regulations on importation of chimpanzees for research. Such a policy is unnecessary and would deepen the plight of an already endangered species.

Clinical effects of alpha-interferon dose variation on laryngeal papillomas.

Abstract: Achieving optimal clinical control of recurrent respiratory papillomatosis with prolonged treatment with human leukocyte (alpha) interferon appears to be dose-related and often requires individualized dosage elevation. Six of eight patients in this study needed a maximum dose of 18 × 106 IU/week for part of the therapy period to achieve better disease control. The strong correlation found between dosage and response suggests that it is the interferon causing the effect on disease expression, not just the unpredictable nature of the disease. An effect on papilloma growth was observed in all patients during IFN therapy. Three patients were able to be tapered off IFN with only minimal recurrence seen in one patient. No toxic side effects were observed.

Translational selection in the expression of the hepatitis B virus envelope proteins.

Abstract: The region of the hepatitis B virus (HBV) genome coding for the viral envelope proteins contains three inphase ATGs that are conserved among viral subtypes. Each of these ATGs can be used as mRNA initiation codons. The three translated proteins share a carboxy-terminal region (the S protein) and extend amino-terminally to include the pre-S2 region in the middle (M) protein, and the pre-S1 and pre-S2 regions in the large (L) protein. We have inserted the HBV DNA coding for the M protein into a baculovirus expression vector. Infected insect cells transcribe a mRNA that is initiated solely within a baculovirus promoter, and that contains the initiator codons for both M and S proteins. Although these cells primarily secrete the M protein, the major translational product is the S protein, which is not secreted. This preferential translation, the result of the use of an internal initiator codon, demonstrates that the regulation of HBV envelope protein production can occur at the translational level.

Extraction of process chemicals from labile biological mixtures with halogenated hydrocarbons

Abstract: A method of removing lipid soluble chemicals from a biological material containing the lipid soluble chemicals comprising bringing the biological material containing the lipid soluble chemicals into contact with an effective amount of an organic alcohol whose solubility in an aqueous solution at the temperature of use is ≦0.6% and/or a halogenated hydrocarbon having more than one type of halogen, agitating the resultant mixture and achieving a phase separation of the biological material and the organic alcohol and/or halogenated hydrocarbon. The method is particularly useful for producing relatively virus free physiologically acceptable plasma.

Inactivation of the Hutchinson strain of hepatitis non-A, non-B virus in intravenous immunoglobulin by β-propiolactone

Abstract: beta-propiolactone (beta-PL) treatment has been evaluated for its ability to inactivate 10(3.5) chimpanzee infectious doses (CID50) of the Hutchinson strain of hepatitis non-A, non-B virus (HNANBV). Two chimpanzees were inoculated with a beta-PL-treated immunoglobulin solution to which this dose of the titrated virus had been added prior to beta-PL treatment. beta-PL treatment was performed in accordance with the production procedure used for a licensed intravenous immunoglobulin preparation. Neither animal developed hepatitis. When subsequently challenged with the same spiked immunoglobulin solution that had not been beta-PL treated, both animals developed clear-cut hepatitis non-A, non-B. The results of this experiment demonstrate that beta-PL treatment is effective for the inactivation of hepatitis non-A, non-B virus in intravenous immunoglobulin.

Antibodies recognizing human serum albumin are not elicited by immunization with preS2 sequences of the hepatitis B virus envelope protein.

Abstract: Antibodies to the preS2 region of the hepatitis B virus (HBV) envelope protein and to human serum albumin (HSA) were allegedly detected at about the same level in sera of humans with acute or chronic hepatitis B [Hellstrom et al., 1986]. It was claimed that anti-HSA arises as a result of an immune response to the preS2 sequence and that it was involved in hepatocellular damage. Over 100 sera from animals and humans immunized with HBsAg containing preS2 sequences, or with synthetic peptides from the preS1, preS2, and S regions of the HBV env protein were assayed for anti-HSA. The results revealed the following: 1) Immunization with the native preS2 sequence or with unconjugated synthetic peptides derived from that sequence does not result in elicitation of anti-HSA. Therefore the alleged appearance of anti-HSA during hepatitis B cannot be directly related to an anti-preS2-specific immune response. 2) Some synthetic peptides, whether or not they were derived from the preS2 sequence, when linked to certain carriers, but not to others, elicited in rabbits an anti-HSA response, which was markedly lower than the response to the homologous peptide. These anti-HSA antibodies could be separated from anti-preS2-specific antibodies by affinity chromatography and did not recognize the synthetic peptide used for immunization. The use in active immunoprophylaxis of hepatitis B of unconjugated peptides from the preS2 sequence with proven high immunogenicity will avoid carrier/linker-mediated induction of antibodies not relevant to protection against HBV.

Blood grouping with the Olympus PK7100 testing system.

Abstract: Summary The Olympus PK7100, an automated microtitre blood typing machine, identifies, aliquots, and dilutes samples, adds reagents, photometrically detects agglutination, and records reaction results, with a throughput of 240 samples/h. A total of 20 147 donors was tested in parallel with the Groupamatic 360. Of these, 207 could not be ABO typed after two runs. Three samples typed as B by the Olympus were found to be A2B. Seventy-seven could not be typed for D after two runs. Of these, 55 were Rh positive and 22 negative. The Olympus identified 37 of 48 Du positive samples as Rh positive, while it typed three as Rh negative, and eight as ‘uncertain’. None of these samples was identified as Rh positive by the Groupamatic 360. The Olympus PK7100 is accurate, reliable, easy to operate, and capable of high throughput with minimal operator intervention.