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C Botting

Publications and source records attributed to C Botting.

8 recordsLinked to original sources
PubMed1996

Construction, purification and immunogenicity of antigen-antibody-LTB complexes.

An oligonucleotide, encoding a short epitope peptide tag, termed Pk, was inserted at the 3'-end of the gene coding B-subunit of Escherichia coli heat-labile enterotoxin (LTB). The presence of the Pk epitope on LTB-Pk was used to construct novel macromolecular assemblies comprising LTB-Pk, an anti-Pk mAb, (mAb SV5-P-k) and Pk-linked recombinant SIV proteins. The 1:1:1 stoichiometry of such complexes was ensured by binding LTB-Pk to one arm of mAb SV5-P-k and an SIV-Pk antigen to the other arm of the antibody. Such SIV-mAb-LTB macromolecular complexes bound to GM1-ganglioside in vitro, and when immunized systemically into mice were highly immunogenic, inducing both humoral and cell-mediated responses to the recombinant SIV antigens.

Amino Acid Sequence↗
PubMed1994

Purification of antibody-antigen complexes containing recombinant SIV proteins: comparison of antigen and antibody-antigen complexes for immune priming.

This paper describes a general procedure for the two-step purification of recombinant proteins as antibody-antigen complexes in which there is no uncomplexed antibody or antigen. In this way, immune complexes containing the p17, p27, vpr and vpx proteins of simian immunodeficiency virus (SIV) have been purified. Antibody-antigen complexes are more immunogenic than antigen when administered either alone or with alum. The significance of the work is that this general method could be modified for the manufacture of immune complexes for incorporation into multivalent vaccines.

Animals↗
PubMed1994

Expression and purification of nonglycosylated SIV proteins, and their use in induction and detection of SIV-specific immune responses.

Two commercially available expression vectors were modified to generate plasmids pGEXcPk and pQ9cPk. Proteins expressed from pGEXcPk and pQ9cPk had a short oligopeptide tag termed Pk at their carboxy termini and either glutathione S-transferase (GST) or a small histidine (His) tag, respectively, at their N termini. GST fusion proteins can be purified on immobilized glutathione and proteins coupled to the His tag selectively bind to Ni(2+)-NTA columns. The Pk tag is recognized by monoclonal antibody (MAb) SV5-P-k, previously produced in our laboratory. Thus proteins expressed from the pGEXcPk and pQ9cPk vectors can be purified in a two-step procedure, first via the N-terminal tag and second via the C-terminal tag. The combination of two affinity purification steps significantly improves the antigen purity and selects for full-size proteins. Moreover, by using the MAbSV5-P-k in the second purification step, Pk-linked antigens can be assembled directly into solid matrix-antibody-antigen (SMAA) complexes for use as vaccines. The genes for nef, endonuclease, p15, p17, p27, protease, Rev, reverse transcriptase (rt), tat, vif, vpr, and vpx of simian immunodeficiency virus (SIV mac 251) were cloned and expressed as both GST-SIV-Pk and His-SIV-Pk proteins. Multivalent SMAA complexes were made that contained His-p17-Pk, His-p27-Pk, His-rt-Pk, His-vpx-Pk, and His-vpr-Pk. Following two immunizations of mice with this mixture, antibodies could be detected to all five SIV antigens. When compared to single-protein immunizations, the immunogenicity of some of the proteins in this cocktail was either enhanced or decreased. Mice were also immunized with His-p17-Pk or His-p17-Pk-antibody complexes in the presence or absence of alum. The antibody-antigen complexes induced two- to four-fold higher antibody levels than antigen alone but did not appear to be more immunogenic in inducing lymphoproliferative responses. Sera from SIV-infected macaques were tested for the presence of antibodies reacting with the recombinant proteins by Western blot analysis. Antibodies to endonuclease, p15, p17, p27, rt, and vif were readily detected, antibodies against protease and vpx were present at much lower levels, but no antibodies were detected to nef, rev, tat, or vpr. Thus, we have developed a comprehensive range of reagents (available on request) that can be used to examine immune responses to SIV in both mice and monkeys.

Animals↗
PubMed1993

Postpolio syndrome in New Zealand: a survey of 700 polio survivors.

AIMS: To examine the experience of postpolio syndrome amongst a group of survivors of polio currently resident in New Zealand. METHODS: A sample of 700 responded to a request for volunteers to take part in a postal survey concerning their experience of polio and postpolio symptoms. RESULTS: The mean age of respondents was 59 years. Two-thirds of them were women. The year of polio infection was between 1915 and 1962 with the majority (54%) being in the 1945-56 period. Most were under 16 years of age (73%) at the time. Paralysis and weakness in limbs and back were the most common symptoms in the acute phase of the infection. Symptoms of the postpolio syndrome were reported by significant numbers. Increasing weakness in muscle functioning in one or more area was evident amongst 38% of the sample. Generalised muscle weakness was reported by 47%, increasing muscle wastage by 17%, difficulty swallowing by 16% and shortness of breath on waking by 10%. Pain in joints was reported by 60% and excessive tiredness by 48%. After controlling for age there was little evidence that the symptoms increased with years since the acute polio infection. CONCLUSIONS: The experience of postpolio symptoms was common amongst this group of polio survivors. It is estimated that there are between 3,000 and 5,000 polio survivors in New Zealand who may be suffering postpolio symptoms. The implications of this for primary and secondary health care provision are discussed.

Adult↗
PubMed1991

Cancer and health screening in Canterbury general practices.

The general practitioners in the Canterbury Area Health Board area were surveyed for their screening policies for cancer and medical conditions. Responses were obtained from 210 (79%), 55% of whom had age/sex registers. Ninety-seven percent provided cervical smears, usually at 1-2 year intervals; 62% offered a female smear taker. Smears were initiated opportunistically by 76%, by age/sex register (47%) or on request by 27%. Breast cancer was screened by 69% using mammography and by 59% using breast physical examination; 73% taught breast self examination. Mammography was recommended every two years for women aged 50-64 years by 45% of responders, and annually to women aged 40-50 years by 19%. Mammography was initiated opportunistically by 88%, on request by 70% and using an age/sex register by 21%. Melanoma was screened by 66%, colorectal cancers in those at high risk by 42%. Testicular self examination was promoted by 43%. Ninety-one percent screened for hypertension, and 51% for hyperlipidaemia, 54% for diabetes mellitus in people without risk factors. Smoking (97%) and alcohol intake (82%) were usually inquired for, and safe sex practices by 59%. Established screening modalities were recommended by most practitioners, but the frequency exceeded current guidelines in many cases; opportunistic screening predominated.

Family Practice↗
PubMed1988

Impact of private medical insurance on the demand for child consultations with general practitioners.

Data were collected on over 4000 consultations with general practitioners for childhood complaints. Almost half of these consultations (46%) were covered by private medical insurance. A larger proportion of consultations for respiratory conditions were covered by insurance (50%) compared to those for nonrespiratory conditions (42% insured). Amongst those with respiratory conditions larger proportions of consultations for upper respiratory tract infections were insured (50%) than was the case for the potentially more serious conditions of asthma or otitis media (46% insured). The delay in seeking consultation for respiratory conditions was examined. In consultations for followup or persisting problems there was less delay among the insured (8.7 days) than amongst the noninsured (11 days). Those consultations initiated by the patient (or parent) were more likely to be covered by insurance (50%) than those which had been initiated by the doctor or other agency (44%). The findings are discussed in terms of the implications for equity and efficiency in health care.

Adolescent↗