Use of a species-specific antibody for demonstrating mouse neurons transplanted to rat brains

ABSTRACT Species-specific antibody epitopes within several major immunoreactive protein orthologs of Ehrlichia species have recently been identified and molecularly characterized. In this study, dominant B-cell epitopes within the acidic (pI 5.35) ankyrin repeat-containing 200-kDa major immunoreactive protein (gp200) of Ehrlichia canis were defined. The E. canis gp200 gene (4,263 bp; 1,421 amino acids) was cloned and expressed as four (N-terminal, 1,107 bp; N-internal, 910 bp; C-internal, 1,000 bp; and C-terminal, 1,280 bp) overlapping recombinant proteins. The N-terminal, C-internal, and C-terminal polypeptides (369, 332, and 426 amino acids, respectively) were strongly recognized by antibody, and the major epitope(s) in these polypeptides was mapped to four polypeptide regions (40 to 70 amino acids). Smaller overlapping recombinant polypeptides (14 to 15 amino acids) spanning these regions identified five strongly immunoreactive species-specific epitopes that exhibited conformational dependence. The majority of the epitopes (four) were located in two strongly acidic (pI 4 to 4.9) domains in the distal N- and C-terminal regions of the protein flanking the centralized ankyrin domain-containing region. The amino acid content of the epitope-containing domains included a high proportion of strongly acidic amino acids (glutamate and aspartate), and these domains appear to have important biophysical properties that influence the antibody response to gp200.

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Species-Specific Antibody Responses to the Recombinant 53-Kilodalton Excretory and Secretory Proteins in Mice Infected with Trichinella spp.

Clinical and Vaccine Immunology ◽

10.1128/cvi.00467-07 ◽

2008 ◽

Vol 15 (3) ◽

pp. 468-473 ◽

Cited By ~ 17

Author(s):

Isao Nagano ◽

Zhiliang Wu ◽

Yuzo Takahashi

Keyword(s):

Escherichia Coli ◽

Recombinant Protein ◽

Recombinant Proteins ◽

Specific Antibody ◽

Expression System ◽

Amino Acid Sequences ◽

Antibody Responses ◽

Secretory Proteins ◽

Muscle Larvae ◽

Species Specific

ABSTRACT The 53-kDa proteins in larval excretory and secretory (E-S) products were expressed from five Trichinella species (T. spiralis, T. britovi, T. nativa, T. pseudospiralis, and T. papuae), using the Escherichia coli expression system, and the antibody responses to the 53-kDa recombinant proteins in mice infected with Trichinella spp. were analyzed by Western blotting. The 53-kDa protein is conserved among the five Trichinella species, with >60% similarity in amino acid sequences. The 53-kDa recombinant proteins of T. spiralis and T. pseudospiralis reacted to sera from mice infected with T. spiralis and T. pseudospiralis at 8 days postinfection (p.i.), respectively. An antibody against the 53-kDa recombinant protein of T. spiralis recognized the 53-kDa protein in the crude extracts from adult worms and 30-day p.i. muscle larvae and E-S products from muscle larvae of T. spiralis but did not recognize any proteins from T. pseudospiralis. The sera from the mice infected with T. spiralis strongly reacted with the 53-kDa recombinant protein of T. spiralis but did not react with the 53-kDa recombinant proteins of T. britovi, T. nativa, T. pseudospiralis, and T. papuae. Similarly, the sera from mice infected with T. britovi, T. nativa, T. pseudospiralis, or T. papuae strongly reacted with the 53-kDa recombinant proteins of T. britovi, T. nativa, T. pseudospiralis, or T. papuae, respectively. These results showed that the 53-kDa recombinant proteins provide early and species-specific antibody responses in mice infected with Trichinella spp.

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Identification of a Glycosylated Ehrlichia canis 19-Kilodalton Major Immunoreactive Protein with a Species-Specific Serine-Rich Glycopeptide Epitope

Infection and Immunity ◽

10.1128/iai.01494-06 ◽

2006 ◽

Vol 75 (1) ◽

pp. 74-82 ◽

Cited By ~ 38

Author(s):

Jere W. McBride ◽

C. Kuyler Doyle ◽

Xiaofeng Zhang ◽

Ana Maria Cardenas ◽

Vsevolod L. Popov ◽

...

Keyword(s):

Amino Acids ◽

Specific Antibody ◽

Tandem Repeats ◽

Ehrlichia Canis ◽

Small Subset ◽

Amino Acid Homology ◽

Amino Terminal ◽

Neutral Sugars ◽

Carboxyl Terminal ◽

Species Specific

ABSTRACT Ehrlichia canis has a small subset of major immunoreactive proteins that includes a 19-kDa protein that elicits an early Ehrlichia-specific antibody response in infected dogs. We report herein the identification and molecular characterization of this highly conserved 19-kDa major immunoreactive glycoprotein (gp19) ortholog of the Ehrlichia chaffeensis variable-length PCR target (VLPT) protein. E. canis gp19 has substantial carboxyl-terminal amino acid homology (59%) with E. chaffeensis VLPT and the same chromosomal location; however, the E. chaffeensis VLPT gene (594 bp) has tandem repeats that are not present in the E. canis gp19 gene (414 bp). Consistent with other ehrlichial glycoproteins, the gp19 protein exhibited a larger-than-predicted mass (∼3 kDa), O-linked glycosylation sites were predicted in an amino-terminal serine/threonine/glutamate (STE)-rich patch (26 amino acids), carbohydrate was detected on the recombinant gp19 protein, and the neutral sugars glucose and galactose were detected on the recombinant amino-terminal polypeptide. E. canis gp19 composition consists of five predominant amino acids, cysteine, glutamate, tyrosine, serine, and threonine, concentrated in the STE-rich patch and a carboxyl-terminal domain predominated by cysteine and tyrosine (55%). The amino-terminal STE-rich patch contained a major species-specific antibody epitope strongly recognized by serum from an E. canis-infected dog. The recombinant glycopeptide epitope was substantially more reactive with antibody than the synthetic (nonglycosylated) peptide, and periodate treatment of the recombinant glycopeptide epitope reduced its immunoreactivity, demonstrating the importance of a carbohydrate immunodeterminant(s). The gp19 protein was present on reticulate and dense-cored cells, and it was found extracellularly in the fibrillar matrix and associated with the morula membrane, the host cell cytoplasm, and the nucleus.

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Assembly and targeting of adaptin chimeras in transfected cells.

The Journal of Cell Biology ◽

10.1083/jcb.123.1.67 ◽

1993 ◽

Vol 123 (1) ◽

pp. 67-77 ◽

Cited By ~ 48

Author(s):

M S Robinson

Keyword(s):

Plasma Membrane ◽

Specific Antibody ◽

The Other ◽

Cytoplasmic Staining ◽

Transfected Cells ◽

Rat Fibroblasts ◽

Species Specific ◽

Terminal Domains

Adaptors are the components of clathrincoated pits and vesicles that attach the clathrin to the membrane. There are two types of adaptors in the cell: one associated with the plasma membrane and one associated with the TGN. Both adaptors are heterotetramers consisting of two adaptins (alpha and beta for the plasma membrane; gamma and beta' for the TGN), plus two smaller proteins. The COOH-terminal domains of the adaptins form appendages that resemble ears, connected by flexible hinges. Unlike the other adaptor components, the COOH termini of the alpha- and gamma-adaptins show no homology with each other, suggesting that they might provide the signal that directs the adaptors to the appropriate membrane. To test this possibility, the COOH-terminal ears were switched between alpha- and gamma-adaptins and were also deleted. All of the constructs contained the bovine gamma-adaptin hinge, enabling them to be detected with a species-specific antibody against this region when transfected into rat fibroblasts. Immunoprecipitation indicated that the engineered adaptins were still fully capable of assembling into adaptor complexes. Immunofluorescence revealed that in spite of their modified ears, the constructs were still able to be recruited onto the appropriate membrane; however, the ear-minus constructs gave increased cytoplasmic staining, and replacing the gamma-adaptin ear with the alpha-adaptin ear caused a small amount of colocalization with endogenous alpha-adaptin in some cells. Thus, the major targeting determinant appears to reside in the adaptor "head," while the ears may stabilize the association of adaptors with the membrane.

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A Case-Control study identifying highly tuberculosis-exposed, HIV-1-infected but persistently TB, tuberculin and IGRA negative persons with M. tuberculosis specific antibodies in Cape Town, South Africa

10.1101/2020.07.07.20147967 ◽

2020 ◽

Author(s):

Elouise E. Kroon ◽

Craig J. Kinnear ◽

Marianna Orlova ◽

Stephanie Fischinger ◽

Sally Shin ◽

...

Keyword(s):

South Africa ◽

T Cell ◽

Specific Antibody ◽

Cape Town ◽

Case Control ◽

Antibody Testing ◽

Specific Antibodies ◽

Antibody Titres ◽

Species Specific ◽

Hiv 1

AbstractBackgroundMycobacterium tuberculosis (Mtb) infection is inferred from positive results of T-cell immune conversion assays measuring Mtb-specific interferon gamma production or tuberculin skin test (TST) reactivity. Certain exposed individuals, termed resisters or early clearers, do not display T-cell immune conversion in these assays. Here we report a hitherto unknown extreme form of this phenotype: HIV-1-infected persistently TB, tuberculin and IGRA negative (HITTIN).MethodsA community-based case-control design was used to systematically screen and identify HIV-1-infected persons (aged 35-60 years) who met stringent study criteria, and follow up for repeat IGRA and TST testing. Participants had no history of TB despite living in TB hyper-endemic environments in Cape Town, South Africa with a provincial incidence of 681/100,000. Mtb-specific antibodies were measured using ELISA and Luminex.FindingsWe identified 48/286(17%) individuals who tested persistently negative for Mtb-specific T-cell immunoreactivity (three negative Quantiferon results and one TST = 0mm) over 206±154 days on average. Of these, 97·2% had documented CD4 counts<200 prior to antiretroviral therapy (ART). They had received ART for 7·0±3·0 years with a latest CD4 count of 505·8±191·4 cells/mm3. All HITTIN sent for further antibody testing (n=38) displayed Mtb-specific antibody titres.InterpretationImmune reconstituted HIV-infected persons can be persistently non-immunoreactive to Mtb, yet develop species-specific antibody responses. Exposure is evidenced by Mtb-specific antibody titers. Identifying individuals displaying such extreme resistance to T-cell immune conversion indicates the importance of early events of TB pathophysiology in the context of HIV-infection that so far have received only scant attention.FundingFunding provided by National Institutes of Health [1R01AI124349-01].

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The Fluorescent Antibody Test in the Serological Diagnosis of the Causative Organisms of Tropical Eosinophilia and Filariasis

Journal of Helminthology ◽

10.1017/s0022149x00027231 ◽

1968 ◽

Vol 42 (1-2) ◽

pp. 57-64 ◽

Cited By ~ 13

Author(s):

L. G. Jayewardene ◽

Y. Wijayaratnam

Keyword(s):

Short Duration ◽

Specific Antibody ◽

Fluorescent Antibody ◽

Antibody Test ◽

Serological Diagnosis ◽

Human Parasite ◽

Fluorescence Reaction ◽

Species Specific ◽

Specific Reactions ◽

Active Disease

The results of the fluorescent antibody test with sera from tropical eosinophilia and filariasis cases tested against four species of microfilariae are reported. The majority of sera from both groups showed the presence of antibody to the human parasite W. bancrofti. In the filarial group only those with clinically active disease of short duration showed a positive fluorescence reaction. Some sera of both groups reacted positively with microfilariae of the animal parasites B. ceylonensis and Setaria sp. These were species specific reactions as seen by diminished or absence of fluorescence after absorption of specific antibody.

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THE INFLUENCE OF HEATING THE IMMUNIZATION MATERIAL UPON THE ANTIBODY-INVOKING EFFECTIVENESS OF THE TYPE-SPECIFIC AND SPECIES-SPECIFIC ANTIGENS OF TYPE II PNEUMOCOCCUS CELLS

Journal of Experimental Medicine ◽

10.1084/jem.47.1.131 ◽

1928 ◽

Vol 47 (1) ◽

pp. 131-150 ◽

Cited By ~ 3

Author(s):

Emidio L. Gaspari ◽

John Y. Sugg ◽

William L. Fleming ◽

James M. Neill

Keyword(s):

Specific Antibody ◽

Experimental Comparison ◽

Type Ii ◽

Specific Antibodies ◽

Comparable Effect ◽

Specific Antigens ◽

Species Specific ◽

Maximum Content ◽

The Ideal

This paper presents an experimental comparison of the effect of heating of the immunization material upon the antibody-invoking effectiveness of the type-specific (SP) and species-specific (P) antigens of Type II pneumococci. Heating of the pneumococcus suspension (vaccine) invariably decreased the production of species-specific antibodies (anti-P) without a comparable effect upon the production of type-specific antibodies (anti-S). For diagnostic typing purposes, the ideal antipneumococcus serum should contain the maximum content of type-specific, and the minimum of species-specific antibody. Our results with forty-one rabbits indicate that the ideal serum from the type-specific standpoint would be obtained by immunization with the heated cells of virulent pneumococci over a comparatively short immunization period; and that the only thing gained by continued immunization or by the use of unheated bacteria at any stage of the immunization, is an increase in the species-specific antibody which is undesireable in sera to be used for diagnostic purposes.

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