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Biomedical subjects

D C Jackson

Publications and source records attributed to D C Jackson.

At least 109 records · Page 6Linked to original sources
PubMed1988

Genetic control and fine specificity of the immune response to a synthetic peptide of influenza virus hemagglutinin.

The immune response to a synthetic peptide, H3 HA1(305-328), representing the C'-terminal 24 amino acid residues of the HA1 chain of the hemagglutinin of the H3 subtype of influenza virus is controlled by genes in the I region of the major histocompatibility complex. Mice of the H-2d haplotype are high responders and produce antibody for several months after a single injection of peptide without carrier. Mice of the H-2b, H-2k, and H-2q haplotypes are low antibody responders. Investigation of recombinant and congenic mouse strains revealed that high responsiveness requires the genes that encode the I-Ed molecule. Immunoassays, involving direct binding to analogs of this peptide and inhibition by both these analogs and synthetic epitopes, were used to analyze the specificity of the polyclonal response. In BALB/c mice, the primary antibody response is directed principally against the antigenic site 314-LKLAT-318, whereas the secondary response after a boost is predominantly directed to a distinct site, 320-MRNVPEKQT-328. The T-cell response to the peptide H3 HA1(305-328), as measured by antigen-induced proliferation of primed T cells in vitro, is also I-Ed restricted in high-responder H-2d mice and is directed against an antigenic site that does not require the four C-terminal residues unique to the H3 influenza subtype. A different epitope appears to be recognized by T cells from CBA (H-2k) mice, which proliferate to a moderate extent on exposure to the peptide but, nevertheless, do not provide help for an antibody response.

Animals↗
PubMed1988

Effect of induced hypercapnia on anaerobic metabolic rate of anoxic musk turtles.

To evaluate the possible effect of induced hypercapnia on anaerobic metabolic rate during anoxia, musk turtles (Sternotherus odoratus) were submerged in N2-equilibrated water at 10 degrees C for 3 days either with (anoxic hypercapnic) or without (anoxic normocapnic) elevated aquatic PCO2 (30-40 Torr). Control animals had access to air at 10 degrees C. Plasma [lactate] was significantly higher (P less than 0.01) in the normocapnic [59.4 +/- 7.4 (SD) mM; n = 22] than in the hypercapnic (47.4 +/- 8.5 mM; n = 19) anoxic turtles, although the hypercapnic turtles had lower blood pH (P less than 0.05). Plasma ion concentrations (Na, K, Cl, Ca, and Mg), however, were no different in the two groups, although all values other than Na were different from control. In some of the animals, [lactate] and [glycogen] (per g wet wt) of skeletal muscle, heart, and liver were measured in addition to blood acid-base values and lactate. Tissue lactates, although significantly elevated from control, and glycogens, although (with the exception of skeletal muscle) significantly reduced from control, were no different in the two anoxic groups. We suggest that these tissue data are more valid indicators of anaerobic metabolic rate than is plasma lactate and therefore conclude that induced hypercapnia does not significantly depress anaerobiosis in musk turtles at 10 degrees C.

Acid-Base Equilibrium↗
PubMed1988

Brain and cerebrospinal fluid ion composition after long-term anoxia in diving turtles.

Prolonged anoxia in turtles is associated with marked disturbances in plasma composition. This study examines brain and cerebrospinal fluid (CSF) ion homeostasis in the freshwater turtle, Chrysemys picta bellii, in response to 8-10 days of submergence anoxia at 10 degrees C. For comparison, it also examines the response to experimental elevation of plasma [K], [Ca], and [Mg] in normoxic turtles. Long-term anoxia resulted in marked changes in brain and CSF composition. These included elevated [K], [Ca], [Mg], and [lactate] and reduced [Cl], and the composition of CSF approached that of plasma. Brain water content increased by 17%, which we suggest was an intracellular edema linked largely to an 11% increase in total brain K. In contrast to the lack of effective homeostasis during anoxia, CSF composition was controlled in normoxic animals. We conclude that there is homeostasis of K, Ca, and Mg in the extracellular fluids of normoxic turtle brain, as in other vertebrates, but that this homeostasis fails during long-term anoxia.

Animals↗
PubMed1987

Direct role of viral hemagglutinin in B-cell mitogenesis by influenza viruses.

The mitogenic activity of influenza virus is a function of the hemagglutinin (HA) molecule. Purified HA is mitogenic for murine B lymphocytes but not T lymphocytes. Furthermore, like the intact virus, HA of the H2 (but not H3) subtype is mitogenic only for B cells expressing the class II major histocompatibility complex glycoprotein I-E. Since virus bearing uncleaved HA is as mitogenic as virus bearing cleaved HA, the membrane fusion activity of the HA molecule is not involved.

Animals↗
PubMed1986

Antigenic determinants of influenza virus hemagglutinin. XII. the epitopes of a synthetic peptide representing the C-terminus of HA1.

A synthetic peptide comprising the C-terminal 24 amino acids of the heavy chain (HA1) of influenza virus hemagglutinin was constructed and examined for antigenic and immunogenic activity. Monoclonal antibodies as well as polyclonal antisera raised against the synthetic peptide were able to bind to intact virus. This binding was greatly enhanced if the virus was first subjected to pH 5, suggesting that this treatment exposes the C-terminus of HA1. Using synthetic analogs of the native sequence it was shown that the epitope recognized by one of the monoclonal antibodies encompasses one or more of the C-terminal four amino acids of HA1 (residues 325-328), which are conserved within subtypes but differ between subtypes, while the other monoclonal antibody recognizes a different epitope which involves at least one of the five variable residues at positions 311-315.

Amino Acid Sequence↗
PubMed1986

Ionic compensation with no renal response to chronic hypercapnia in chrysemys picta bellii.

The ionic compensatory response to CO2 breathing for 3 days was studied on intact and cystectomized turtles at 10 and 20 degrees C. Arterial blood gases, pH, ionized calcium, and the plasma concentrations of Na+, K+, Cl-, total Ca2+, and total Mg2+ were measured periodically. At 20 degrees C, ureteral urine was also collected from bladderless turtles and was analyzed for pH, ions, NH3+, total CO2, osmolality, and titratable acid. When CO2 was breathed there was a compensatory change in the strong-ion difference as manifest by an increase in plasma [HCO3-] that was approximately 10 meq/l both in the 10 and 20 degrees C turtles. The only significant associated strong-ion changes observed consistent with the ionic compensatory response were increases in total and ionized Ca2+ and total Mg2+. These results were unaffected at either temperature by surgical removal of the urinary bladder. Urine collected from cystectomized turtles showed no compensatory increase in acid excretion during hypercapnia; in fact, changes occurred in the opposite direction. Urinary excretion of HCO3- and urine pH increased significantly, whereas titratable acidity decreased significantly. No significant change occurred in ammonia excretion over the three days of hypercapnia. These data argue against compensatory roles for the kidneys and urinary bladder in this species and point to internal ionic exchanges involving bone and shell.

Acid-Base Equilibrium↗
PubMed1985

Antigenic determinants of influenza virus hemagglutinin. XI. Conformational changes detected by monoclonal antibodies.

At pH 5 influenza virus hemagglutinin undergoes an irreversible conformational change (J.J. Skehel, P. M. Bayley, E. B. Brown, S. R. Martin, M. D. Waterfield, J. M. White, I. A. Wilson, and D. C. Wiley (1982). Proc. Natl. Acad. Sci. USA 79, 968-972) which parallels the appearance of fusion activity of this molecule. This paper describes experiments which explore the conformational change using a panel of monoclonal antibodies which define four of the major antigenic sites of this protein. The results indicate that three of the major antigenic sites of hemagglutinin undergo changes when exposed to acid pH. These changes have little effect on the binding avidity of influenza virus to glycophorin, the major receptor present on the red blood cell surface. These findings have been used to postulate a mechanism where the molecule flexes around a central region resulting in rearrangement in space of its component domains on exposure to low pH.

Antibodies, Monoclonal↗
PubMed1985

Conformational changes in influenza virus haemagglutinin and its monomer detected by monoclonal antibodies.

Exposure of influenza virus haemagglutinin to pH 5 results in conformational changes occurring in the molecule which are accompanied by antigenic modifications. Furthermore, isolated haemagglutinin (HA) at a concentration of 0.1 nM undergoes dissociation from the trimeric to a monomeric form when exposed to pH 5. Whether present on intact virus or as the isolated monomer, each form of haemagglutinin from pH 5 exhibits similar alterations in antigenic characteristics. These forms of HA show modifications in the antigenic sites located in the hinge (site C), tip (site B) and subunit interface (site D) regions. Whereas binding of monoclonal antibodies recognizing the tip and interface is abrogated or diminished, binding of antibodies to the hinge region is greatly enhanced following exposure of virus or the monomeric form of HA to pH 5.

Antibodies, Monoclonal↗
PubMed1985

Antibodies elicited by influenza virus hemagglutinin fail to bind to synthetic peptides representing putative antigenic sites.

A number of peptides of the hemagglutinin (HA) of X-31 influenza virus have been synthesised. The amino acid sequences of some of these peptides represent regions of HA which have been postulated [Wiley et al., Nature, Lond. 289, 373-378 (1981)] to form the antigenic sites of this molecule. Animals were immunized with free peptide or peptide conjugated to a carrier and the resulting antisera examined for their capacities to bind to homologous peptide, whole HA, reduced and alkylated HA, and intact virus. Not all peptides examined in this way were immunogenic. Only antibodies raised against the C-terminus of HA1 peptide displayed binding to virus. This antiserum bound to the intact HA but not to the reduced and alkylated form of the molecule. These results raise questions as to the feasibility of using synthetic peptides of the influenza HA in short linear sequences to elicit neutralising antibody.

Amino Acid Sequence↗
PubMed1985

Antigenic determinants of influenza virus haemagglutinin. X. A comparison of the physical and antigenic properties of monomeric and trimeric forms.

Haemagglutinin prepared from influenza virus A/Memphis/1/71 by bromelain digestion was centrifuged through continuous sucrose gradients buffered at pH 7.4 or pH 4.9. From these gradients were isolated two forms of the protein which displayed different equilibrium sedimentation properties. One species behaved as a molecule with a mol. wt. of 190 000, the other with a mol. wt. of 70 000. These results are consistent with the separation of trimeric and monomeric haemagglutinin. A comparison of their antigenic properties, using monoclonal antibodies raised against intact virus, showed that major antigenic differences occur between the two forms of haemagglutinin. None of the monoclonal antibodies reacted with haemagglutinin denatured by reduction and alkylation.

Animals↗
PubMed1985

Central chemical control of ventilation and response of turtles to inspired CO2.

The role of central chemosensors in the overall ventilatory response of freshwater turtles (Chrysemys scripta elegans) to the addition of CO2 in inspired gas was measured. Centrally mediated ventilatory responses were isolated in the unanesthetized animal by combining CO2 breathing and brain ventricular perfusion with mock cerebrospinal fluid (CSF) of varying acid-base status. Breathing 4.5% CO2 resulted in increases in both ventilatory frequency (f) and tidal volume (VT), with increases in VT providing most of the overall ventilatory change. Alterations in the acid-base status of the perfusate produced highly significant changes in f. VT changes were divorced from the acid-base status of the mock CSF perfusate. We therefore conclude that ventilatory changes in turtles, mediated by central chemosensors, are primarily affected by alterations in f. VT changes, associated with acid-base homeostatic mechanisms, are mediated by receptors outside the blood-brain barrier in these animals. On the basis of these data, we hypothesize that the increase in f observed when turtles breathe 4.5% CO2 is primarily mediated by the central chemosensors.

Acid-Base Equilibrium↗
PubMed1983

Changes in the antigenicity of the hemagglutinin molecule of H3 influenza virus at acidic pH.

In order to determine the location and biological significance of the acid-induced conformational change in influenza virus hemagglutinin (HA) reported by Skehel et al., monoclonal antibodies were prepared to the molecule before and after treatment at pH 5.0. These antibodies together with monoclonal antibodies to the different antigenic regions of the H3 HA were used in immunoprecipitation and ELISA binding studies to show that antigenic changes accompanied the conformational change in the HA. Treatment at pH 5.2 or less exposed new determinants on the HA while two antigenic regions, located at the tip and interface of the molecule at neutral pH, were lost or modified. Antigenic sites in the loop and hinge regions defined by the available monoclonal antibodies were not affected by the conformational change. Monoclonal antibodies specific for the acid-induced conformation efficiently inhibited hemagglutination of the virus at low pH but were extremely poor inhibitors of virus-induced red blood cell hemolysis at its pH optimum of 5.1. These antibodies were unable to neutralize viral infectivity under neutral or acidic conditions. Antibodies specific for the non-acid-treated HA conformation failed to inhibit hemagglutination at low pH values but were able to both inhibit hemolysis of red blood cells and neutralize virus infectivity. Residual unmodified HA after pH 5.0 treatment could explain the inhibition of hemolysis and infectivity by monoclonal antibodies in each of the different antigenic areas.

Animals↗
PubMed1983

Chemical and antigenic characterization of the carbohydrate side chains of an Asian (N2) influenza virus neuraminidase.

The Pronase-released neuraminidase heads from the Asian influenza virus A/Tokyo/3/67 contain four oligosaccharide units attached at asparagine residues 86, 146, 200, and 234. Chemical analysis of the isolated tryptic, chymotryptic, or thermolytic glycopeptides shows that the oligosaccharide side chains attached at residues 86 and 200 are essentially of the oligomannoside (simple or Type II) variety containing two residues of N-acetylglucosamine, five residues of mannose, and less than molar ratios of galactose and fucose. The carbohydrate side chains attached at residues 146 and 234 are of the N-acetyllactosamine (complex or Type I) type and contain N-acetylglucosamine, mannose, galactose, and fucose. The complex oligosaccharide unit at residue 146 is unusual in that it also contains N-acetylgalactosamine, a sugar residue rarely found in N-glycosidically linked carbohydrates. Antigenic analysis of these four isolated glycopeptides showed that only the N-acetyllactosamine oligosaccharide unit at asparagine residue 146 was capable of binding to antibodies raised against uninfected chick chorioallantoic membranes and is hence antigenically related to chick embryo host antigen.

Amino Sugars↗
PubMed1983

Intracellular and extracellular acid-base and electrolyte status of submerged anoxic turtles at 3 degrees C.

Specimens of fresh water turtles (Chrysemys picta bellii) were acclimated to 3 degrees C and then submerged in completely anoxic water for time periods of up to 12 weeks. Blood withdrawn via indwelling arterial catheters was analysed for plasma pH, PCO2, bicarbonate concentration, [lactate], [Na+], [K+], [Ca2+] and [Mg2+], and tissue samples of skeletal muscle, liver and cardiac muscle were excised. Samples of skeletal muscle were analysed for intracellular pH (DMO), [lactate], [Na+], [K+], [Ca2+], and [Mg2+], and samples of liver and cardiac muscle for intracellular pH and [lactate] during normoxia and after 1, 2, 4, 8 and 12 weeks of anoxia. Arterial plasma pH fell from 8.0 during normoxia to lower than 7.2 concomitant with a reduction in plasma [HCO3-] after 12 weeks of anoxia due to the production of large amounts of lactic acid. The intracellular pH (pHi) of heart muscle and liver dropped in parallel or even more than plasma pH, whereas pHi in skeletal muscle changed less resulting in a delta pHi/delta pHe value of less than 0.6. Intracellular [lactate] and [Ca2+] increased considerably, but attained concentrations much smaller than those observed in the extracellular compartment. The intracellular concentrations of K+, Na+ and Mg2+ were also significantly affected, the changes, however, were small in comparison with those observed for Ca2+ and lactate concentration. The water distribution between intra- and extracellular compartments remained essentially unaffected by anoxia. It is concluded that the considerable increase in extracellular Mg2+ and Ca2+ cannot be the result of release from muscle cells and has to be attributed to release from skeleton and shell.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid-Base Equilibrium↗
PubMed1983

The equilibrium constant for the interaction between a monoclonal Fab fragment and an influenza virus neuraminidase.

The affinity or equilibrium constant between an Fab fragment derived from monoclonal IgG directed against influenza virus neuraminidase was measured as 4.1 X 10(7) M-1. The method, which makes use of an air-driven ultracentrifuge, is simple and uses extremely small amounts (10(-11) mol) of material. Furthermore, interpretation of the data is based on sound theoretical considerations. The technique also allows m.w. of the interacting species to be measured and the stoichiometry of the reaction to be determined.

Animals↗
PubMed1982

Plasma ion balance of submerged anoxic turtles at 3 degrees C: the role of calcium lactate formation.

Freshwater turtles, Chrysemys picta bellii, were submerged in groups of 7 at 3 degrees C in O2-free water for 1, 2, 4, 8 and 12 weeks. Blood samples from these turtles and from 10 normoxic turtles at 3 degrees C were analyzed for plasma concentrations of lactic acid, total CO2, Na+, K+, Cl-, Ca2+, total calcium, total magnesium and osmolality. Total lactate rose during anoxia to a mean peak value of 145 mM, but the decrease in HCO-3 and Cl- and increase in K+ balanced less than 40% of the lactate. Total calcium and total magnesium rose respectively by 9.5 and 6.0 times the normoxic values after 12 weeks, at which time free [Ca2+] was 25.0 mEq (37% of the total calcium). To evaluate the possible role of bound calcium in ion balance, test solutions with calcium, but with and without 145 mM lactate, were tested for free Ca2+. In the presence of lactate, over two-thirds of the total calcium combined with lactate- to form a calcium lactate complex (possibly CaLactate+). Based on these data, it is concluded that most of the bound plasma calcium in the anoxic turtles was combined with lactate. By assuming that magnesium reacts similarly with lactate, a complete account of plasma ion balance is accomplished and the turtle's plasma ionic response to extreme lactic acidosis is described. Plasma osmolality increased during anoxia by 100 mOsm and matched the mM rise in total measured and calculated ions.

Animals↗