1. THE EFFECT OF NICOTINE ON PRIMARY HUMORAL IMMUNE RESPONSE IN MICE Mentors:AP SinisaRistic Authors: NenadTodorovic MD Vladimir TuruntasSnezana Milosevic StevanBasurovic Ivan Zivkovic Boris Pejic

2. Introduction Aim Method Results Conclusion Quantitatively, nicotine represents the co most important alcaloide of tobacco. Primarily, it comes into our organism by masticating tobacco, or inhaling tobacco smoke. When we intake nicotine in our organism, 90% of it metabolises quickly ( half-life of about 120 minutes), whereas 10% of it is distributed in unchanged condition to tissues and body liquids.

3. IntroductionAim Method Results Conclusion Nicotine expresses its direct features by means of nicotine cholinergic receptors (nACh) whose endogenous ligand is acetylcholine. Research significance of the influence of small nicotine dosages on humoral immune response in experimental animals is seen in possibility of their analogy with endogenic cholinergic regulation of immune response in animals and men.

4. IntroductionAim Method Results Conclusion Asymptomatically active smokers have more leucocytes in peripheral blood in comparison with non-smokers. Immunomodulatory, namely immunotoxic effects of cigarette smoke are obtained after exposing experimental animals to particulate phase cigarette smoke. After mice’s inhaling cigarette smoke in the period of 5 days, there is an increase in cells number which produces antibodies in the spleen, but not in the neck and mediastinale lymph nodules.

5. Introduction Aim Method Results Conclusion Our aim was to examine whether nicotine causes changes of intensity of humoral immune response in mouse which was applied subcutaneously in dosages of 0.5, 1 and 2 mg per weight.

6. Introduction AimMethod Results Conclusion Our experiments were carried out on mice, an Albino type, which were 4-6 weeks old. All animals in our experiments were males. Experiments were carried out on mice weighing 18-22 grams.

7. Introduction Aim Method Results Conclusion The animals were randomly divided into control and experimental group. Nicotine was subcutaneously applied to experimental animals in the period of five days, after they had been immunised with sheep erithrocytes. According to the nicotine dosage which was used, the animals were divided into three groups: A:0,5 mg/kg per weight daily; B: 1, 0 mg/kg per weight daily; C: 2, 0 mg/kg per weight daily;

8. Introduction Aim Method Results Conclusion . The humoral immune response in this resaerch was quantified by direct method of hemolitic plaques– “Plaque forming cell assay – PFC’’, AFC experiment (antibody forming cell). The spleen suspension was obtained by disintigration of spleen pulp resection by means of fine curve pipette in 4 ml of Hanks solution and then pressing this suspension through plastic syringe, volume of 5ml without a needle five times , and then through syringe and needle No 2 five times more. After determining the number of spleen cells by means of hematimeter, their number was fitted to 3 x 106per ml in Hanks solution.

9. Introduction Aim Method Results Conclusion Figure 1: Descriptive statistics of antybody-productive cells (PFC)/ 106 of splecocytes in mice treated with different nicotine dosages (0.5mg, 1 mg and 2 mg per day) which was applied in the period of 5 days after immunisation.

10. Introduction Aim Method ResultsConclusion Figure 2: Results of Mann-Whitney comparison of number of antibody-productive cells (PGC)/ 106 of splecocytes in mice treated with different nicotine dosages (0.5mg, 1 mg and 2 mg per day) which was applied in the period of 5 days after immunisation.

11. Introduction Aim Method ResultsConclusion Graph 1: Median number of antibody-productive cells/106 of Msplenocytesin mice treated with different nicotine dosages (0.5mg, 1 mg and 2 mg/kg) which was applied in the period of 5 days after immunisation.