ABSTRACT
A monoclonal antibody (MAb) was produced by immunization of a BALB/c mouse with a conjugated morphine C6-hemisuccinated derivative (MHS) to cationized bovine serum albumin (cBSA). The hybridoma clones were screened by indirect ELISA using MHS-BSA. The best hybridoma clone was subcloned thrice by limiting dilution. This hybridoma was found to be of IgG2b class and subclass and contained lambda light chain. The affinity of the MAb to morphine was obtained 2.8×10(9) M(-1). The titer of the cell culture supernatant was at least 1:800. The MAb was cross-reacted with codeine (100%) and apomorphine (16.5%), but not with heroin, naloxone, naltrexone, or papaverine. Morphine was conjugated to HRP using a mixed anhydride method and a direct competitive ELISA was designed using anti-morphine MAb. The assay was sensitive over the 50 ng/mL to 5 µg/mL concentration range. In conclusion, this MAb is useful for the development of immunoassays to measure morphine in urine.
Subject(s)
Antibodies, Monoclonal/immunology , Morphine/immunology , Animals , Antibody Affinity/immunology , Apomorphine/immunology , Codeine/immunology , Cross Reactions/immunology , Enzyme-Linked Immunosorbent Assay/methods , Hybridomas/immunology , Immunization/methods , Immunoassay/methods , Immunoglobulin G/immunology , Mice , Mice, Inbred BALB CABSTRACT
The alkaloid oxoglaucine reduced CD4+ cell clones in adult mice and decreased CD4+, CD8+ and Ig+ levels in newborn mice. It prevented the increase of CD8+ and Ig+ clones induced by Candida albicans (C. albicans) in adult mice. TNF-alpha serum accumulation was inhibited by oxoglaucine in C. albicans infection and adjuvant arthritis. Treatment with oxoglaucine of arthritic mice, followed by inoculation with C. albicans enhanced the host resistance against the pathogen.
Subject(s)
Adjuvants, Immunologic/therapeutic use , Apomorphine/analogs & derivatives , Arthritis, Experimental/complications , Candida albicans/drug effects , Candidiasis/complications , Animals , Apomorphine/immunology , Apomorphine/therapeutic use , Biological Assay , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Candidiasis/drug therapy , Candidiasis/immunology , Female , Flow Cytometry , Mice , Tumor Necrosis Factor-alpha/analysis , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
It has been established that injection of apomorphine leads to the dose-dependent enhancement of the immune response in CBA mice whereas that of haloperidol suppresses the number of rosette-forming cells. After thymectomy apomorphine does not stimulate rosette-formation. This indicates that the thymus participates in the realization of apomorphine action. Preliminary injection of haloperidol prevents the apomorphine-induced enhancement of the immune response. The results suggest that the enhancement is mediated through the activation of dopamine receptors.
