1. PRESENTED BYPRESENTED BY:-:-
Prafulla C TiwariPrafulla C Tiwari
M.Pharm (2M.Pharm (2ndnd
sem)sem)
PharmacologyPharmacology
UNDER THE
GUIDANCE OF:-
MR. RUPESH GAUTAM
(Lecturer)
Department Of
Pharmacology

2.  The immune system evolved to discriminate self
from nonself.
 Multicellular organism shows immune response by
destroying infectious invaders (microbes) while
leaving normal cells intact.
 Immunity system shows two types of responses
that are-
1.Innate immunity
2.Adaptive immunity

3. 1.Innate, or natural immunity-It is primitive, does
not require priming, and have relatively low affinity,
but is broadly reactive. The major effectors of innate
immunity are granulocytes,
monocytes/macrophages, natural killer cells and
mast cells.
2. Adaptive, or learned, immunity-It is antigen
specific, depends upon antigen exposure and have
very high affinity. The major effectors of adaptive
immunity are B and T lymphocytes. B lymphocytes
make antibodies; T lymphocytes function as helper,
cytolytic cells.

4.  Immunomodulators are the agents that modulates
the immune system by suppress or stimulate the
immune response.So, these are also divided into two
parts that are-
1.Immunosuppressants &
2.Immunostimulants

5. 1.IMMUNOSUPPRESSANTS-
Immunosuppressive drugs are used to
dampen the immune response in organ
transplantation and autoimmune disease.
(a)Specific T-cell inhibitors-Cyclosporine,Tacrolimus
(b)Cytotoxic drugs-Azathioprine, Cyclophosphamide,
Methotrexate, Chlorambucil, Mycophenolate
mofetil(MMF)
(c)Glucocorticoids-Prednisolone
(d)Antibodies-Muromonab CD3, Antithymocyte
globulin(ATG), Rho(D) immunoglobulin

6. 2.IMMUNOSTIMULANT-
These drugs are used to stimulate the immune response
in case of immunodeficiency.
(a) Levamisole (ergamisole)
(b) Thalidomide (thalomid)
(c) Bacillus Calmette-Guerin (BCG)
(d) Interferons- gamma1b & beta1a
(e) Interleukin-2

7. Screening method
 Acute systemic anaphylaxis in rats
 Anti-anaphylactic activity (schultz-dale reaction)
 Passive cutaneous anaphylaxis
 Arthus type immediate hypersensitivity
 Delayed type hypersensitivity
 Reversed passive arthus reaction
 Adjuvant arthritis in rats

8. Acute systemic anaphylaxis in rats
 Strain: 10-20 Female Sprague-Dawley rat(120g)
 Firstly immunized by i.m. injection of 10mg/kg
highly purified ovalbumin.
 Simultaneously 1ml of Bordetella pertusis
suspension injected intraperitoneally.
 After 11 days animals injected with i.v.
injection of 25mg/kg highly purified
ovalbumin
 18hr prior to challenge;
test – dexamethasone 1-10mg/kg s.c.
control-vehicle

9.  Evaluation-
after treatment compared treated and control
group for their shock symptoms and mortality
counted.
Statistical calculation is performed using the
chi-square test.

10. Modification-
 By Davis and Evans(1973)-This experiment have
also been performed in guinea pigs and in mice.
Anaphylactic bronchospasm can be measured in
isolated guinea pig lungs.
 By Ufkes and Ottenhof(1984)- studied on Brown-
Norway rats
Given a suspension of trinitrophenyl haptenized
ovalbumin together with AlPO4.
Bronchial and cardiovascular function were
studied after treatment with antiallergic agents and
antigen challenge.

11.  Elwood et al.(1992)- studied on Brown-Norway
rats.
They studied the effect of dexamethasone and
cyclosporin A on hyperresponsiveness and
inflammatory cell responses.

12. Anti-anaphylactic activity
(Schultz-Dale reaction)
 Animal-guinea pig of both sex
 Body wt.-300-350g
 Sensitized with alum precipitated egg
albumin.
 Give injection of 0.125ml of egg
albumin in each foot pad and 0.5ml
subcutaneously.

13.  After 4 weeks animals are killed and the ileum
is dissected out.
 About 2-3cm long are mounted in an organ
bath containing Tyrode solution at 370
C.
 Contractility of the ileum strips is tested by
adding standardin one organ bath and test
compound in another organ bath.

14. Biochemical estimation-
 Carbon clearance test- for phagocytic response
Phagocytic index=K(sample)/K(control)
 Hemagglutinating antibody titer

15. Evalution-
The contractions are recorded by a polygraph.
Modification-
 This method modified by testing histamine
release in the lung after challenging with egg
albumin.
 Koppel et al.(1981) developed a method to
induce contraction of mouse trachea by
antigen.
 Omote et al.(1994) modified method by using
sensitized guinea pigs.

16. Passive cutaneous anaphylaxis
 Animal-male rats
 Body wt.-100g
Principle-
 Formation of antigen-antibody complex induces the
release of mediator from mast cells.This results
increase in permeability of the vessel walls and
leakage of plasma.

17. Procedure-
 Antiserum are injected intradermally in to the
shaved dorsal skin of rats.
 After 24hr each animal is challenged with the
intravenous administration of 0.1ml of 2.5% Evans
blue dye containing 25mg/ml of egg albumin.
 Test compound is also administer along with the
antigen.
 After 30 min.,animals are sacrificed.
 Amount of Evans blue dye that leaked at the site of
reaction is extracted and determined
colorimetrically at 620milimicron wavelength.

18. Evaluation-
 Amount of Evans blue that extracted from passive
cutaneous anaphylactic reaction of control group is
compared with the treated group.
Modification-
 Goose and Blair(1969)- used Bordetella pertusis as
antigen in passive cutaneous anaphylaxis
experiments in the rat.
 Patterson et al.(1971)- tested passive cutaneous
reactivity to antihuman IgE in rhesus monkey.

19. Arthus type immediate hypersensitivity
 Animal-rats of both sex
 Strain-Wistar or Sprague-Dawley
 Body wt.-220-280g
Principle-
 Antigen-antibody induced reaction leading to an
inflammatory factors that characterised by
edeme,hamorrhage and vasculitis.

20. Procedure-
 Seven days prior to experiment rats are sensitized
by i.m. administration of 0.5ml of the ovalalbumin
suspension.
 1st
group(treated group)- 1hr prior test compound are
administered and challenged with 0.1ml of
ovalalbumin in left hind paw
 2nd
group(positive control)-sensitized animals treated
with solvent alone.
 3rd
group(negative control)-nonsensitized animals
treated with solvent.

21. Evalution-
 The change in footpad thickness is expressed as
percent change from the vehicle control group.
Thickness can be measured by calipers.
Modification-
 Omote et al.(1994)-sheep red blood cell suspension
used for immunization.

22. Delayed type hypersensitivity
Principle-
 Delayed type hypersensitivity is a reaction of cell
mediated immunity and becomes visible after 16-
24hr.
Procedure-
 7days prior,rats are sensitised by i.m. administration
of 0.5ml ovalbumin.
 After 7 days again 0.1ml of 0.04% of ovalbumin
injected in the left hind paw.

23.  Footpad thickness is measured immediately and
24hr after of administration.
Modification-
 Mizukoshi et al.(1994)-They use sheep red blood
cells for immunization.

24. References-
 Vogel.H.G. , second edition, Analgesics, page
no:797-801.
 Bigonia P. & Rana A.C. “Immunomodulatory
activity of EUPHORBIA NERIIFOLIA leaf
alcoholic extract on rats” Indian drugs, feb.2008