scholarly journals Biased Immunoglobulin G1 Isotype Responses Induced in Cattle with DNA Expressing msp1a of Anaplasma marginale

1999 ◽  
Vol 67 (7) ◽  
pp. 3481-3487 ◽  
Author(s):  
Appudurai Arulkanthan ◽  
Wendy C. Brown ◽  
Travis C. McGuire ◽  
Donald P. Knowles
Keyword(s):  
Immune Responses ◽  
Surface Protein ◽  
Anaplasma Marginale ◽  
Surface Proteins ◽  
Dependent Manner ◽  
Genes Encoding ◽  
Major Surface Proteins ◽  
Major Surface ◽  
Dose Dependent ◽  
Homologous Challenge

ABSTRACT Immunization with the native major surface protein 1 (MSP1) (a heterodimer containing disulfide and noncovalently bonded polypeptides designated MSP1a and MSP1b) of the erythrocytic stage ofAnaplasma marginale conferred protection against homologous challenge (G. H. Palmer, A. F. Barbet, W. C. Davis, and T. C. McGuire, Science 231:1299–1302, 1986). The MSP1a polypeptide possesses a conserved neutralization-sensitive epitope. In the present study, the immune response to DNA-mediated immunization using msp1a was studied. The plasmid pVCL/MSP1a, which encodes the complete msp1a gene of A. marginaleunder the control of human cytomegalovirus immediate-early enhancer/promoter and intron A, was constructed. The immune responses elicited by immunization with pVCL/MSP1a into cardiotoxin-induced regenerating muscle were evaluated in mice and cattle. Antibody reactive with native MSP1a was detected in pooled sera of immunized BALB/c mice 3 weeks following primary immunization. Two calves seronegative for A. marginale were immunized four times, at weeks 0, 3, 7, and 13, with pVCL/MSP1a. By 8 weeks, both calves responded to MSP1a with an antibody titer of 1:100, which peaked at 1:1,600 and 1:800 by 16 weeks after the initial immunization. Interestingly, immunoblotting with anti-immunoglobulin G1 (anti-IgG1) and anti-IgG2 specific monoclonal antibodies revealed a restricted IgG1 anti-MSP1a response in both animals. T-lymphocyte lines, established after the fourth immunization, proliferated specifically againstA. marginale homogenate and purified MSP1 in a dose-dependent manner. These data provide a basis for an immunization strategy to direct bovine immune responses by using DNA vaccine vectors containing single or multiple genes encoding major surface proteins ofA. marginale.

scholarly journals Upregulation of Intercellular Adhesion Molecule 1 and Proinflammatory Cytokines by the Major Surface Proteins of Treponema maltophilum and Treponema lecithinolyticum, the Phylogenetic Group IV Oral Spirochetes Associated with Periodontitis and Endodontic Infections

2005 ◽  
Vol 73 (1) ◽  
pp. 268-276 ◽  
Author(s):  
Sung-Hoon Lee ◽  
Kack-Kyun Kim ◽  
Bong-Kyu Choi
Keyword(s):  
Proinflammatory Cytokines ◽  
Group Iv ◽  
Surface Proteins ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Pdl Cells ◽  
Genes Encoding ◽  
Major Surface Proteins ◽  
Endodontic Infections ◽  
Major Surface

ABSTRACT Treponema maltophilum and Treponema lecithinolyticum belong to the group IV oral spirochetes and are associated with endodontic infections, as well as periodontitis. Recently, the genes encoding the major surface proteins (Msps) of these bacteria (MspA and MspTL, respectively) were cloned and sequenced. The amino acid sequences of these proteins showed significant similarity. In this study we analyzed the functional role of these homologous proteins in human monocytic THP-1 cells and primary cultured periodontal ligament (PDL) cells using recombinant proteins. The complete genes encoding MspA and MspTL without the signal sequence were cloned into Escherichia coli by using the expression vector pQE-30. Fusion proteins tagged with N-terminal hexahistidine (recombinant MspA [rMspA] and rMspTL) were obtained, and any possible contamination of the recombinant proteins with E. coli endotoxin was removed by using polymyxin B-agarose. Flow cytometry showed that rMspA and rMspTL upregulated the expression of intercellular adhesion molecule 1 (ICAM-1) in both THP-1 and PDL cells. Expression of proinflammatory cytokines, such as interleukin-6 (IL-6) and IL-8, was also induced significantly in both cell types by the Msps, as determined by reverse transcription-PCR and an enzyme-linked immunosorbent assay, whereas IL-1β synthesis could be detected only in the THP-1 cells. The upregulation of ICAM-1, IL-6, and IL-8 was completely inhibited by pretreating the cells with an NF-κB activation inhibitor, l-1-tosylamido-2-phenylethyl chloromethyl ketone. This suggests involvement of NF-κB activation. The increased ICAM-1 and IL-8 expression in the THP-1 cells obtained with rMsps was not inhibited in the presence of the IL-1 receptor antagonist (IL-1ra), a natural inhibitor of IL-1. Our results show that the Msps of the group IV oral spirochetes may play an important role in amplifying the local immune response by continuous inflammatory cell recruitment and retention at an infection site by stimulation of expression of ICAM-1 and proinflammatory cytokines.

Anaplasma marginale(Rickettsiales: Anaplasmataceae): recent advances in defining host–pathogen adaptations of a tick-borne rickettsia

Parasitology ◽  
2004 ◽  
Vol 129 (S1) ◽  
pp. S285-S300 ◽  
Author(s):  
K. M. KOCAN ◽  
J. DE LA FUENTE ◽  
E. F. BLOUIN ◽  
J. C. GARCIA-GARCIA
Keyword(s):  
Antigenic Variation ◽  
Control Strategies ◽  
Anaplasma Marginale ◽  
Surface Proteins ◽  
Host Cells ◽  
Developmental Cycle ◽  
Persistent Infections ◽  
Phylogenetic Studies ◽  
Major Surface Proteins ◽  
Major Surface

The tick-borne intracellular pathogenAnaplasma marginale(Rickettsiales: Anaplasmataceae) develops persistent infections in cattle and tick hosts. While erythrocytes appear to be the only site of infection in cattle,A. marginaleundergoes a complex developmental cycle in ticks and transmission occurs via salivary glands during feeding. Many geographic isolates occur that vary in genotype, antigenic composition, morphology and infectivity for ticks. In this chapter we review recent research on the host–vector–pathogen interactions ofA. marginale. Major surface proteins (MSPs) play a crucial role in the interaction ofA. marginalewith host cells. The MSP1a protein, which is an adhesin for bovine erythrocytes and tick cells, is differentially regulated and affects infection and transmission ofA. marginalebyDermacentorspp. ticks. MSP2 undergoes antigenic variation and selection in cattle and ticks, and contributes to the maintenance of persistent infections. Phylogenetic studies ofA. marginalegeographic isolates usingmsp4andmsp1α provide information about the biogeography and evolution ofA. marginale:msp1α genotypes evolve under positive selection pressure. Isolates ofA. marginaleare maintained by independent transmission events and a mechanism of infection exclusion in cattle and ticks allows for only the infection of one isolate per animal. Prospects for development of control strategies by use of pathogen and tick-derived antigens are discussed. TheA. marginale/vector/host studies described herein could serve as a model for research on other tick-borne rickettsiae.

scholarly journals Development of a Multivalent ISCOM Vaccine against Anaplasmosis

1993 ◽  
Author(s):  
Guy H. Palmer ◽  
Eugene Pipano ◽  
Terry F. McElwain ◽  
Varda Shkap ◽  
Donald P. Knowles, Jr.
Keyword(s):  
T Lymphocytes ◽  
Outer Membrane ◽  
Membrane Surface ◽  
Surface Protein ◽  
The United States ◽  
Surface Proteins ◽  
Efficient Production ◽  
Major Surface ◽  
Homologous Challenge

Anaplasmosis is an arthropod+borne disease of cattle caused by the rickettsia Anaplasma marginale and an impediment to efficient production of healthy livestock in both Israel and the United States. Our research focuses on development of a recombinant membrane surface protein (MSP) immunogen to replace current vaccines derived from the blood of infected cattle. The risk of widespread transmission of both known and newly emergent pathogens has prevented licensure of live blood-based vaccines in the U.S. and is a major concern for their continued use in Israel. Briefly, we accomplished the following in our BARD supported research: i) characterization of the intramolecular and intermolecular relationships of the native Major Surface Proteins (MSP) in the outer membrane; ii) expression, purification, and epitope characterization of the recombinant MSP-2, MSP-3, MSP-4, and MSP-5 proteins required to construct the recombinant ISCOM; iii) demonstration that the outer membrane-Quil A induces CD4+ T lymphocytes specific for the outer membrane polypeptides; iv) identification of CD4+ T lymphocytes that recognize outer membrane polypeptide epitopes conserved among other wise antigenically distinct strains; v) determination that immunization with the outer membrane-Quil A construct does not affect the ability of ticks to acquire or transmit A. marginale; and vi) demonstration that the outer membrane-Quil A construct induces complete protection against rickettsemia upon homologous challenge and significant protection against challenge with antigenically distinct strains, including tick transmission. Importantly, the level of protection against homologous challenge in the MSP vaccinates was comparable to that induced by live blood-based vaccines and demonstrates that development of a new generation of vaccines is feasible.

scholarly journals Conformational Dependence ofAnaplasma marginale Major Surface Protein 5 Surface-Exposed B-Cell Epitopes

1998 ◽  
Vol 66 (6) ◽  
pp. 2619-2624 ◽  
Author(s):  
Devere Munodzana ◽  
Terry F. McElwain ◽  
Donald P. Knowles ◽  
Guy H. Palmer
Keyword(s):  
Surface Protein ◽  
Covalent Modification ◽  
Surface Proteins ◽  
Immunodominant Epitope ◽  
Disulfide Bonding ◽  
Amino Terminal ◽  
Major Surface Proteins ◽  
Major Surface ◽  
Carboxy Terminal ◽  
Epitope Binding

ABSTRACT The Anaplasma marginale outer membrane is composed of immunogenic major surface proteins (MSPs) linked both covalently and noncovalently in multimeric complexes (M. C. Vidotto, T. C. McGuire, T. F. McElwain, G. H. Palmer, and D. P. Knowles, Infect. Immun. 62:2940–2946). Consequently, effective induction of antibody against surface-exposed MSP epitopes has been postulated to require maintenance of MSP secondary through quatenary structures. Using MSP5 as a model and the approach of epitope mapping with recombinant expressed full-length and truncated proteins, we demonstrated that the immunodominant surface epitope bound by monoclonal antibody (MAb) ANAF16C1 required disparate amino- and carboxy-terminal regions of MSP5, indicating the conformational dependence of this epitope. The required amino-terminal MSP5 region included the cysteines involved in intramolecular disulfide bonding. The dependence of the immunodominant epitope on disulfide bonding was confirmed by loss of MAb ANAF16C1 binding to MSP5 following disulfide bond reduction and covalent modification of the reduced sulfhydryl groups. The recognition of the MSP5 immunodominant epitope by antibody induced by protective immunization with A. marginale outer membranes was also conformationally dependent, as shown by the loss of epitope binding following serum adsorption with native but not reduced and denaturedA. marginale. Importantly, the antibody response to all immunodominant MSP5 surface epitopes was restricted to conformationally dependent epitopes, since the binding of polyclonal anti-MSP5 antibody to the A. marginale surface could be blocked by adsorption with native but not denatured and reduced MSP5. These results confirm the importance of the secondary and tertiary structures of MSP epitopes as immune system targets and support the testing of immunogens which maintain the required conformation.

scholarly journals Thermostability of Reovirus Disassembly Intermediates (ISVPs) Correlates with Genetic, Biochemical, and Thermodynamic Properties of Major Surface Protein μ1

2002 ◽  
Vol 76 (3) ◽  
pp. 1051-1061 ◽  
Author(s):  
Jason K. Middleton ◽  
Tonya F. Severson ◽  
Kartik Chandran ◽  
Anne Lynn Gillian ◽  
John Yin ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Thermal Inactivation ◽  
Surface Protein ◽  
Surface Proteins ◽  
Major Surface Protein ◽  
First Order ◽  
Major Surface Proteins ◽  
The Difference ◽  
Major Surface ◽  
Type 3

ABSTRACT Kinetic analyses of infectivity loss during thermal inactivation of reovirus particles revealed substantial differences between virions and infectious subvirion particles (ISVPs), as well as between the ISVPs of reoviruses type 1 Lang (T1L) and type 3 Dearing (T3D). The difference in thermal inactivation of T1L and T3D ISVPs was attributed to the major surface protein μ1 by genetic analyses with reassortant viruses and recoated cores. Irreversible conformational changes in ISVP-bound μ1 were shown to accompany thermal inactivation. The thermal inactivation of ISVPs approximated first-order kinetics over a range of temperatures, permitting the use of Arrhenius plots to estimate activation enthalpies and entropies that account for the different behaviors of T1L and T3D. An effect similar to enthalpy-entropy compensation was additionally noted for the ISVPs of these two isolates. Kinetic analyses with other ISVP-like particles, including ISVPs of a previously reported thermostable mutant, provided further insights into the role of μ1 as a determinant of thermostability. Intact virions, which contain ς3 bound to μ1 as their major surface proteins, exhibited greater thermostability than ISVPs and underwent thermal inactivation with kinetics that deviated from first order, suggesting a role for ς3 in both these properties. The distinct inactivation behaviors of ISVPs are consistent with their role as an essential intermediate in reovirus entry.

scholarly journals Intermolecular relationships of major surface proteins of Anaplasma marginale.

1994 ◽  
Vol 62 (7) ◽  
pp. 2940-2946 ◽  
Author(s):  
M C Vidotto ◽  
T C McGuire ◽  
T F McElwain ◽  
G H Palmer ◽  
D P Knowles
Keyword(s):  
Anaplasma Marginale ◽  
Surface Proteins ◽  
Major Surface Proteins ◽  
Major Surface

scholarly journals A liver-specific nuclear factor interacts with the promoter region of the large surface protein gene of human hepatitis B virus.

1989 ◽  
Vol 9 (11) ◽  
pp. 5189-5197 ◽  
Author(s):  
H K Chang ◽  
B Y Wang ◽  
C H Yuh ◽  
C L Wei ◽  
L P Ting
Keyword(s):  
Hepatitis B Virus ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Surface Protein ◽  
Surface Proteins ◽  
Nuclear Factor ◽  
Hepatoma Cell Lines ◽  
B Virus ◽  
Major Surface Proteins ◽  
Major Surface

The outer envelope of the 42-nm virion of the human hepatitis B virus (HBV) is composed of the large, the middle, and the major surface proteins. Whereas the middle and the major surface proteins are transcribed from the SPII promoter of the pre-S/S gene, the large surface protein is transcribed from the SPI promoter located upstream of SPII. We have previously shown that transcription of SPI (comprising nucleotides [nt] -380 to +17) occurs preferentially in differentiated hepatoma cell lines (H.K. Chang and L.P. Ting, Virology 170:176-183, 1989). In this report, we further demonstrated that a sequence of 95 base pairs in the upstream region of SPI (nt -95 to +17) was necessary and sufficient for such preferential expression in differentiated hepatoma cells. By analysis of the expression of the chloramphenicol acetyltransferase gene in a series of mutants with deletions at the 5' end of SPI, we identified a positive transcriptional cis-acting element mapping at nt -95 to -72 which appears to play a key role in the regulation of the expression of the large surface protein. This region shared a high degree of sequence homology with regulatory sequences of several liver-specific genes from human, mouse, and rat, with a consensus sequence (G/A)GTTA(A/C)TNNT(C/T)NNC(A/C). We further identified a nuclear factor present in the nuclear extracts of differentiated human hepatoma cell lines which interacted specifically with this element of the SPI promoter. This nuclear factor was similar to the rat liver-specific factor HNF-1, since an oligonucleotide containing the recognition sequence of HNF-1 could efficiently compete for the human factor in a footprinting assay. The sequence at nt -93 to -68 which was bound by this factor in SPI was termed the HNF-1-binding element. Activation of the SPI promoter by human differentiated hepatocyte nuclear factor 1, described in this report, probably explains, first, the formation of the 42-nm virion specifically in liver but not in several other tissues despite the synthesis of the middle and the major surface proteins in those tissues, and second, why only differentiated hepatoma cell lines are able to produce 42-nm-like virion particles on transfection by HBV DNA.

A liver-specific nuclear factor interacts with the promoter region of the large surface protein gene of human hepatitis B virus

1989 ◽  
Vol 9 (11) ◽  
pp. 5189-5197
Author(s):  
H K Chang ◽  
B Y Wang ◽  
C H Yuh ◽  
C L Wei ◽  
L P Ting
Keyword(s):  
Hepatitis B Virus ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Surface Protein ◽  
Surface Proteins ◽  
Nuclear Factor ◽  
Hepatoma Cell Lines ◽  
B Virus ◽  
Major Surface Proteins ◽  
Major Surface

The outer envelope of the 42-nm virion of the human hepatitis B virus (HBV) is composed of the large, the middle, and the major surface proteins. Whereas the middle and the major surface proteins are transcribed from the SPII promoter of the pre-S/S gene, the large surface protein is transcribed from the SPI promoter located upstream of SPII. We have previously shown that transcription of SPI (comprising nucleotides [nt] -380 to +17) occurs preferentially in differentiated hepatoma cell lines (H.K. Chang and L.P. Ting, Virology 170:176-183, 1989). In this report, we further demonstrated that a sequence of 95 base pairs in the upstream region of SPI (nt -95 to +17) was necessary and sufficient for such preferential expression in differentiated hepatoma cells. By analysis of the expression of the chloramphenicol acetyltransferase gene in a series of mutants with deletions at the 5' end of SPI, we identified a positive transcriptional cis-acting element mapping at nt -95 to -72 which appears to play a key role in the regulation of the expression of the large surface protein. This region shared a high degree of sequence homology with regulatory sequences of several liver-specific genes from human, mouse, and rat, with a consensus sequence (G/A)GTTA(A/C)TNNT(C/T)NNC(A/C). We further identified a nuclear factor present in the nuclear extracts of differentiated human hepatoma cell lines which interacted specifically with this element of the SPI promoter. This nuclear factor was similar to the rat liver-specific factor HNF-1, since an oligonucleotide containing the recognition sequence of HNF-1 could efficiently compete for the human factor in a footprinting assay. The sequence at nt -93 to -68 which was bound by this factor in SPI was termed the HNF-1-binding element. Activation of the SPI promoter by human differentiated hepatocyte nuclear factor 1, described in this report, probably explains, first, the formation of the 42-nm virion specifically in liver but not in several other tissues despite the synthesis of the middle and the major surface proteins in those tissues, and second, why only differentiated hepatoma cell lines are able to produce 42-nm-like virion particles on transfection by HBV DNA.

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