scholarly journals Keyhole limpet haemocyanin - a model antigen for human immunotoxicological studies

2014 ◽  
Vol 78 (5) ◽  
pp. 1135-1142 ◽  
Author(s):  
Ashwin Swaminathan ◽  
Robyn M. Lucas ◽  
Keith Dear ◽  
Anthony J. McMichael
Keyword(s):  
Keyhole Limpet Haemocyanin

HAPTEN-RADIOIMMUNOASSAY OF PLASMA OESTRADIOL

1973 ◽  
Vol 72 (2) ◽  
pp. 330-344 ◽  
Author(s):  
Peter Doerr
Keyword(s):  
Adult Male ◽  
Normal Values ◽  
Normal Adult ◽  
Adult Males ◽  
Reagent Blank ◽  
Keyhole Limpet Haemocyanin ◽  
Ether Extraction ◽  
The Mean ◽  
Assay Precision ◽  
Plasma Oestradiol

ABSTRACT A hapten-radioimmunoassay for plasma oestradiol is described and information about the reliability of the method is given in detail. Oestradiol-3-hemisuccinate coupled to keyhole limpet haemocyanin is used for immunization of rabbits. The antiserum utilized for the assay is characterized by its titer, affinity and specificity. Following ether extraction and NaOH-light petroleum partition oestradiol is separated from crossreacting oestrogens by TLC. Oxidation of oestradiol on the plate is prevented by mercaptoethanol. To separate free and antibody bound ligand 250 μg dextran-coated charcoal per tube is used in the presence of bovine serum gammaglobulin (1 mg/ml). The between-assay precision based on 15 different determinations of control samples from normal adult male plasma was 9.4% (C. V.). The mean reagent blank value of 31 determinations was equivalent to 0.3 pg oestradiol and the detection limit in terms of the 99% confidence limit for a single blank value, was equivalent to 4.3 pg oestradiol. A procedure for detecting plasma blanks is described. Plasma oestradiol is separated from approximately all concomitant substances originally present in the sample by enzymatic conversion into oestrone and a second TLC. No plasma blanks could be detected with respect to normal adult male plasma. Normal values for adult males based on 51 subjects were characterized by a median of 17.2 pg/ml and the 95 percentiles of 9.5–27.6.

scholarly journals Evaluation of tests based on the antibody response to keyhole limpet haemocyanin and soluble egg antigen to differentiate acute and chronic human schistosomiasis mansoni

2004 ◽  
Vol 99 (suppl 1) ◽  
pp. 97-98 ◽  
Author(s):  
Lílian Beck ◽  
Daniele SM Van-Lüme ◽  
Joelma R de Souza ◽  
Clarice N Lins de Morais ◽  
Wlademir G Melo ◽  
...  
Keyword(s):  
Antibody Response ◽  
Keyhole Limpet Haemocyanin ◽  
Schistosomiasis Mansoni ◽  
Human Schistosomiasis ◽  
Egg Antigen

scholarly journals Desmosomal glycoproteins 2 and 3 (desmocollins) show N-terminal similarity to calcium-dependent cell-cell adhesion molecules

1990 ◽  
Vol 97 (2) ◽  
pp. 239-246
Author(s):  
J.L. Holton ◽  
T.P. Kenny ◽  
P.K. Legan ◽  
J.E. Collins ◽  
J.N. Keen ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Adhesion ◽  
Intercellular Space ◽  
Solid Phase ◽  
Rabbit Antiserum ◽  
High Titre ◽  
Polyclonal Antiserum ◽  
Keyhole Limpet Haemocyanin ◽  
Calcium Dependent ◽  
Dependent Cell

The N-terminal sequence of a mixture of desmosomal glycoproteins 2 and 3 (dg2/3, desmocollins) from bovine nasal epidermis, prepared by electro-elution from polyacrylamide gels, was determined by solid-phase Edman degradation. A sequence of 23 amino acids was obtained. This showed 43% identity with that of the N terminus of the calcium-dependent cell adhesion molecule, N-cadherin. A lesser degree of identity with other members of the cadherin-uvomorulin-L-CAM family was also found. In order to confirm that the sequence was derived from the dg2/3 molecules a rabbit antiserum was raised against a synthetic peptide corresponding to the sequence, conjugated to keyhole limpet haemocyanin (KLH). The antiserum obtained showed high (titre) activity against both the peptide and KLH in ELISA. Each activity could be specifically adsorbed with the appropriate ligand. The antiserum reacted specifically with both dg2 and dg3 of bovine nasal epidermis on immunoblots, this binding was blocked by the N-terminal peptide but was unaffected by KLH. The identity of dg2 and -3 in these preparations was confirmed by immunoblotting with two monoclonal antibodies and one polyclonal antiserum raised against the whole molecules. The N-terminal peptide antiserum was shown to bind to the intercellular space of desmosome profiles by immunoelectron microscopy on ultra-thin frozen sections. One of the two monoclonal antibodies (07–4D) also reacted with the desmosomal intercellular space. dg2 and -3 were shown by Staphylococcus aureus V8 protease digestion to have identical one-dimensional peptide maps. Both the N-terminal antiserum and 07–4D reacted with a V8 fragment of 19,000 Mr derived from dg2 and dg3.(ABSTRACT TRUNCATED AT 250 WORDS)

The behavioural, endocrine and immune responses of suckled calves to weaning and creep-feeding

1995 ◽  
Vol 1995 ◽  
pp. 200-200
Author(s):  
A.M. Mackenzie ◽  
T.G. Rowan ◽  
S.D. Carter ◽  
J.B. Dixon ◽  
J. Tebble
Keyword(s):  
Pilot Study ◽  
Immune Responses ◽  
Lymphocyte Transformation ◽  
Domestic Animals ◽  
Lymphocyte Transformation Test ◽  
Creep Feeding ◽  
Keyhole Limpet Haemocyanin ◽  
Humoral Responses ◽  
Immune Changes ◽  
The Relationship

The effects of husbandry conditions on the immune responses of suckled and bucket-reared calves has previously been reported by Mackenzie et al. (1993 a; 1993b; 1994) and there was a trend for weaning to result in decreased lymphocyte transformation test responses to mitogens along with increased humoral responses to the antigen keyhole limpet haemocyanin (KLH). Mean worker have made measurements of behavioural, endocrine and immune responses to attempt to determine the welfare status of domestic animals. However, there have been few studies where attempts have been made to measure all three parameters together. This pilot study was designed to investigate the relationship between husbandry conditions which may result in behavioural, endocrine and immune changes and to consider possible relationships in these parameters.

Diagnosis of schistosomiasis: antibody detection, with notes on parasitological and antigen detection methods

Parasitology ◽  
1999 ◽  
Vol 117 (7) ◽  
pp. 41-57 ◽  
Author(s):  
J. V. HAMILTON ◽  
M. KLINKERT ◽  
M. J. DOENHOFF
Keyword(s):  
Recombinant Dna ◽  
Infection Intensity ◽  
Public Health Problem ◽  
Antigen Detection ◽  
Detection Methods ◽  
Antibody Detection ◽  
Native Form ◽  
Keyhole Limpet Haemocyanin ◽  
Advantages And Disadvantages ◽  
Wide Range

Schistosomiasis remains a serious world-wide public health problem with a still unfulfilled need for routine cost-effective methods of diagnosis. Such methods are required not only for people in endemic areas, but increasingly for tourists who may have become infected during visits to such places. This article reviews the wide range of immunoassays and antigenic preparations that have been shown to have potential for diagnosis of schistosomiasis by the indirect method of antibody detection. Antigens in native form derived from cercariae, adult worms and eggs are considered, as well as schistosome antigens produced by recombinant DNA technology and the schistosome cross-reactive antigen, keyhole limpet haemocyanin (KLH). Respective advantages and disadvantages of antibody detection, circulating antigen detection and parasitological methods of diagnosis are analysed. It is suggested that due to the relative insensitivity of both parasitology and antigen detection, antibody detection methods could find increasing use in situations of low infection intensity.

Limb injuries, immune response and growth performance of early-weaned pigs in different housing systems

2000 ◽  
Vol 70 (1) ◽  
pp. 73-83 ◽  
Author(s):  
H. R. C. Kelly ◽  
J. M. Bruce ◽  
S. A. Edwards ◽  
P. R. English ◽  
V. R. Fowler
Keyword(s):  
Immune Response ◽  
Live Weight ◽  
Immune Challenge ◽  
Food Conversion ◽  
Weaned Pigs ◽  
Foot Injuries ◽  
Keyhole Limpet Haemocyanin ◽  
Housing Systems ◽  
Expanded Metal ◽  
Limb Injuries

AbstractThe welfare of early-weaned pigs in different housing systems was examined using a range of indicators. Three replicates of 20 pigs were weaned at 6·4 kg live weight into either: (a) deep-straw; (b) Straw-Flow®; (c) large flatdeck; (d) small flatdeck. A kenneled lying area was provided in (a) and (b). The floor in (c) and (d) was expanded metal. Stocking densities were 0·23 m2 per pig in (a), (b) and (c) and 0·17 m2 per pig in (d). After 4 to 5 weeks (19·6 kg live weight), 16 pigs from each pen were moved into Straw-Flow grower pens (0·68 m2 per pig) and observed until slaughter at 90·6 kg. The remaining pigs were grouped together to examine the effects of mixing. At weaning, 24·1% of pigs had foot injuries. Pigs with foot injuries recovered quickly on deep-straw but the injuries increased in flatdecks immediately after weaning (Straw-Flow was intermediate). Injuries were more persistent in the small flatdeck. Deep-straw prevented adventitious bursitis of the hock. After 1 month in the grower pens, there was no residual treatment effect. Antibody response was examined by immune challenge with keyhole limpet haemocyanin (KLH). Immunoglobulin (Ig) response was greater in straw pens. Pigs mixed at 20 kg showed a decreased immune response to KLH challenge. Growth and food intake showed no treatment effects; food conversion ratio was poorer in (c) than in (b) or (d).

The behavioural, endocrine and immune responses of suckled calves to weaning and creep-feeding

1995 ◽  
Vol 1995 ◽  
pp. 200-200
Author(s):  
A.M. Mackenzie ◽  
T.G. Rowan ◽  
S.D. Carter ◽  
J.B. Dixon ◽  
J. Tebble
Keyword(s):  
Pilot Study ◽  
Immune Responses ◽  
Lymphocyte Transformation ◽  
Domestic Animals ◽  
Lymphocyte Transformation Test ◽  
Creep Feeding ◽  
Keyhole Limpet Haemocyanin ◽  
Humoral Responses ◽  
Immune Changes ◽  
The Relationship

The effects of husbandry conditions on the immune responses of suckled and bucket-reared calves has previously been reported by Mackenzie et al. (1993 a; 1993b; 1994) and there was a trend for weaning to result in decreased lymphocyte transformation test responses to mitogens along with increased humoral responses to the antigen keyhole limpet haemocyanin (KLH). Mean worker have made measurements of behavioural, endocrine and immune responses to attempt to determine the welfare status of domestic animals. However, there have been few studies where attempts have been made to measure all three parameters together. This pilot study was designed to investigate the relationship between husbandry conditions which may result in behavioural, endocrine and immune changes and to consider possible relationships in these parameters.

The rôle of cortisol and β-endorphin in the response of the immune system to weaning in lambs

1998 ◽  
Vol 66 (2) ◽  
pp. 397-402 ◽  
Author(s):  
S. M. Rhind ◽  
H. W. Reid ◽  
S. R. McMillen ◽  
G. Palmarini
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Stress Hormone ◽  
Keyhole Limpet Haemocyanin ◽  
Weaning Stress ◽  
Cell Mediated Immune Response ◽  
Acth Treatment ◽  
Induced Changes ◽  
The Relationship

AbstractThe relationship between weaning stress-induced changes in stress hormone profiles and immune function was investigated in groups of 10 lambs immunized against adrenocorticotrophic hormone (ACTH; treatment A) or fi-endorphin (treatment B) to reduce the circulating concentrations of cortisol and fi-endorphin respectively. Control animals (treatment C) were immunized against a porcine thyroglobulin carrier protein. Application of weaning stress was associated with significantly elevated plasma cortisol concentrations but no significant increase in fi-endorphin concentrations in C lambs. Immunization against ACTH suppressed the post-weaning increase in cortisol concentration. This was associated with a transient reduction in the lymphocyte stimulation response to keyhole limpet haemocyanin (KLH) antigen in the A animals but there was no effect on the antibody response or interferon-y production by antigen stimulated lymphocytes. There were no significant effects of immunization against fi-endorphin on the capacity to mount antibody or cell-mediated immune responses. It is concluded that weaning stress-induced increases in cortisol did not inhibit the immune response. Since cortisol concentrations and the cell mediated immune response at 8 days after immunization were positively associated it is concluded that these indices are not independent measures of stress.

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