Using anti-adipocyte Using anti-adipocyte monoclonal antiboies for monoclonal antiboies for

controlling the controlling the development of adipose development of adipose

tissue.tissue.

Dr. Ziad JaradatDr. Ziad Jaradat

Polyclonal antibodies have been raised Polyclonal antibodies have been raised against fat cells or adipocyte plasma against fat cells or adipocyte plasma membrane proteins of rats, rabbits, membrane proteins of rats, rabbits, pigs… etc. studies in rats showed that pigs… etc. studies in rats showed that injection of anti-adipocyte polyclonal injection of anti-adipocyte polyclonal antisera resulted in long term reduction antisera resulted in long term reduction of the number of adipocyte in internal of the number of adipocyte in internal fat depots probably due to adipocyte fat depots probably due to adipocyte lysis.lysis.

A decrease in total body fat as well as of A decrease in total body fat as well as of dorsal and perirenal adipose tissues was dorsal and perirenal adipose tissues was obtained in experimental rabbits. In sheep, obtained in experimental rabbits. In sheep, several fat depots (i.e backfat, several fat depots (i.e backfat, subcutaneous, omental (subcutaneous, omental (One of the folds of the peritoneum that connect the stomach with other abdominal organs) , perirenal , perirenal and kidney pelvic) were reduced after and kidney pelvic) were reduced after injection of antisera against adipocyte injection of antisera against adipocyte plasma membranes.plasma membranes.

However, these experiments did not However, these experiments did not show out obvious improvement of show out obvious improvement of carcass production. In contrast, in carcass production. In contrast, in porcine such anti-sera led to a long porcine such anti-sera led to a long term fat reduction and to an increase term fat reduction and to an increase in lean carcass development.in lean carcass development.

Drawback;Drawback; the anti-sera is not the anti-sera is not specific, so absorption with extracts specific, so absorption with extracts of none adipose tissue is the method of none adipose tissue is the method to increase their specificity, however, to increase their specificity, however, this method is laborious and this method is laborious and expensive.expensive.

Therefore, a monoclonal antibody Therefore, a monoclonal antibody would be the ultimate goal. A highly would be the ultimate goal. A highly specific monoclonal antibody for fat specific monoclonal antibody for fat cell surface proteins has been cell surface proteins has been produced. produced.

ExperimentalExperimental Treatment of young piglets with anti-Treatment of young piglets with anti-

adipocyte monoclonal antibody.adipocyte monoclonal antibody.

Monoclonal antibody preparation was Monoclonal antibody preparation was injected intraperitoneally in young large injected intraperitoneally in young large white pigs two and 5 days after birth. Pigs white pigs two and 5 days after birth. Pigs were sacrificed 38 days after birth, and were sacrificed 38 days after birth, and various tissues were analyzed. Heart, spleen various tissues were analyzed. Heart, spleen and liver were not affected, whearase all the and liver were not affected, whearase all the adipose tissue were slightly decreased in adipose tissue were slightly decreased in weight.weight.

The back fat and subcutaneous ham The back fat and subcutaneous ham adipose tissue reached the most adipose tissue reached the most significant reductions. In addition, significant reductions. In addition, adipocyte were small in size than the adipocyte were small in size than the control one. Finally the total amount of control one. Finally the total amount of lipid in the carcass was significantly lipid in the carcass was significantly decreased without affecting other decreased without affecting other organs or tissues.organs or tissues.

When the monoclonal antibody was When the monoclonal antibody was injected into five day old piglets and injected into five day old piglets and were left to reach 50-100 kg body were left to reach 50-100 kg body weight, it appeared that the weight, it appeared that the subcutaneous layer in the pigs was less subcutaneous layer in the pigs was less as well as the adipose tissue in the as well as the adipose tissue in the shoulder, back fat and the kidney area shoulder, back fat and the kidney area without affecting the gross carcass without affecting the gross carcass weight.weight.

Immunocastration of farm Immunocastration of farm animalsanimals

Traditional castration generates stress to the Traditional castration generates stress to the animals and decreasing their efficiency. animals and decreasing their efficiency. Therefore, it is important to find a none-invasive Therefore, it is important to find a none-invasive alternative. alternative.

Immunization against sex hormones such as Immunization against sex hormones such as LHRH, LH, FSH, and HCG can affect reproduction LHRH, LH, FSH, and HCG can affect reproduction therefore, these hormones were the target of therefore, these hormones were the target of immunocastration. Immunization against immunocastration. Immunization against gonadal steroids, ova and sperm were also gonadal steroids, ova and sperm were also considered.considered.

Immunization against Immunization against LHRHLHRH

LHRH is the first intracellular LHRH is the first intracellular messenger of the control of fertility. messenger of the control of fertility. Immunological neutralization of Immunological neutralization of LHRH would expect to block pituitary LHRH would expect to block pituitary secretion of LH and FSH and will lead secretion of LH and FSH and will lead to gonadal quiescence.to gonadal quiescence.

In lambs, the immunnutralisation In lambs, the immunnutralisation reduced testis weight and the reduced testis weight and the concentration of testosterone in serum concentration of testosterone in serum which also reduced sexual behavior. which also reduced sexual behavior.

Likewise, immunization of heifers Likewise, immunization of heifers against LHRH lead to reduction of against LHRH lead to reduction of fertility equivalent to that of fertility equivalent to that of ovariectomized heifer. ovariectomized heifer.

However the effect of this vaccine is However the effect of this vaccine is reversible and subsequent fertility was reversible and subsequent fertility was normal. A vaccine against LHRH is normal. A vaccine against LHRH is commercialized under the name commercialized under the name VaxstrateVaxstrate and is available in Australia. and is available in Australia. What happens is that the administration What happens is that the administration of the vaccine suppresses ovarian of the vaccine suppresses ovarian cyclicity in post pubertal females for a cyclicity in post pubertal females for a period of six to eight months.period of six to eight months.

After this time the effect of the vaccine After this time the effect of the vaccine disappears and the ovarian function disappears and the ovarian function returns to normal. To make the vaccine returns to normal. To make the vaccine more efficient, it could be administered more efficient, it could be administered with adjuvants which prolong its effect.with adjuvants which prolong its effect.

Another vaccine in the market is called Another vaccine in the market is called ImprovacImprovac, which was used mainly in , which was used mainly in pigs also produced positive results and pigs also produced positive results and pigs treated with this vaccine grew more pigs treated with this vaccine grew more rapidly producing heavy boars with rapidly producing heavy boars with improved meat quality through the improved meat quality through the prevention and control of boar taint prevention and control of boar taint (spoilage and diseases).(spoilage and diseases).

Immunization against Immunization against gonadal steroidsgonadal steroids

Anti-progestrone monoclonal antibody Anti-progestrone monoclonal antibody completely blocked pregnancy. The completely blocked pregnancy. The action of antibody in females was action of antibody in females was associated with a failure to initiate an associated with a failure to initiate an implantation response. The most implantation response. The most pronounced effect, however, was a pronounced effect, however, was a break of embryonic development at a break of embryonic development at a stage prior to cavitation.stage prior to cavitation.

Passive immunization against progesterone Passive immunization against progesterone shortly after mating interferes with early shortly after mating interferes with early hormone dependent steps which are essential hormone dependent steps which are essential for normal embryonic development. for normal embryonic development.

In contrast, immunization against testosterone In contrast, immunization against testosterone was not characterized by inhibition of sexual was not characterized by inhibition of sexual behavior in rams and there was no suppression behavior in rams and there was no suppression of testicular function. of testicular function.

Sperm and Ova antigensSperm and Ova antigens

Antibodies made against sperm or ova Antibodies made against sperm or ova will block gamete interactions required will block gamete interactions required for fertilization. Zona pelluida forms a for fertilization. Zona pelluida forms a coating around mammalian eggs. coating around mammalian eggs.

The ZP is comprised essentially of three The ZP is comprised essentially of three major glycoproteins which are involved major glycoproteins which are involved in sperm binding during their in sperm binding during their penetration through the zona. penetration through the zona.

ZP3 was identified as a major sperm ZP3 was identified as a major sperm receptor as was targeted as most likely receptor as was targeted as most likely female gamete antigen for a female gamete antigen for a contraceptive vaccine. In several contraceptive vaccine. In several species immunization of females species immunization of females against ZP antigens can lead to control against ZP antigens can lead to control of fertility.of fertility.

Now sperm antigens can be used to Now sperm antigens can be used to immunize males or females while ZP immunize males or females while ZP antigens are effective only in females.antigens are effective only in females.

Thank youThank you