Practical Class 2

Immunology
Sef Lucr. Dr Adriana Muntean
Hemograma

• automaticaly

(video cell )
CBC
 CBC
- anemia
Hb, HCT,(H)
- polycythemie

- leucocytosis
Leucocyte
- leucopenia
- Cells anomalies

-thrombocytosis
Thrombocytes
-thrombocytopenia
 Normal values

WBC 5,000 – 10,000 (11.000) /cube.mm.

PMN 40-75 %

L 20-50 %

Eo 1-6 %

Mo 1-6 %

B 1 %
 Anomalies of Leucocytes

leucocytosis leucopenia
neutrophilia neutropenia
limphocytosis lymphopenia
eosinophilia
bazophilia
monocitosis
leukemia
 Neutrophils
• Infections/inflammation
• Tumoral
• Mieloprolipherative diseases:
– Thrombocitaemia/Mielofibrosis
– Increase Hct or Thrombocytes; splenomegaly;
adenopathies
– Chronic myeloid Leukemia
• Corticosteroids treatment
 Neutropenia
• Benigne-etnical (Afro-Caribbean).
• Infections- viruses, -elders
• Drugs/recent chemotherapy
• Haematological diseases (Hb; thrombocytes).
• Megaloblastic Anemia (Hb/MCV).
• Immune:
– Autoimmune: SLE, RA (sd. Felty- adds the
splenomegaly).
 Eosinophilia
Frequent Rare
• Drugs • Conective tisues
• Allergies diseases
– Asthma; eczema; • Skin autoimmune
urticari diseases:
• Parasites – Pemphigus;
infestations • Tumors:
– Hodgkin disease
• Sarcoidosis

Eosinophil
 Limfocitosis
• Infections bacterial/viruses
• Limphoprolipherative diseases:
– CLL
– Limphoma
– Hairy cells Leukemia
 Limphopenia
• SLE/RA
• Viruses: HIV
• Chemoterapy
• Tumors – de ex. Boala Hodgkin
• Sarcoidosis
• Find viruses leading to lymphopenia
 Flowcytometry

• Optical characterization of different particles.

• Identify and sorts the cells due to surface antigens.
• Is using fluorescent compounds

+
Mark
Incubation
 Electronics

Detectori de lumină
Partea
de fluide

FL2 PE
Computer
FL1 FITC

Lumină SSC

laser
488nm FSC
 Presentation of the data

.
Surface markers detected
 ANTIGEN

What is an Antigen?

• Any substance (usually foreign) that binds specifically to an

antibody, or a T cell receptor, is referred to as an ‘antigen’.
• A substance capable of inducing a specific immune response is
called an antigen.
• Immunogenicity and antigenicity are the two attributes of the
antigen.
• A microorganism may have more Ags on the surface

• Antibody A
Antigen binding site (Paratope)

Antigens:
Com ponents o f c ell w all
( A n antigen)
Bacterial cell

Antibody B

Antigenic determinants
on antigen ( Epitopes)
 ANTIBODIES
Immunoglobulins
Electroforesis of serum (video immelec and radial)
 Antigen/Ab detection techniques

Immunofluorescence (IF)- autoantibodies

(video fluoresc)

Immunocapture- ELISA – tumors

markers, Ab (video Elisa, RADAS)

Polymerase chain reaction (PCR) – viruses,

bacteria, tumors (video PCR)
• Examples of autoantibody used in
autoimmune diseases diagnosis

• What viruses can we determine by PCR?

 Tumors markers
– Solubiles RIA / ELISA:
PSA-prostate cancer
– Celular
 Complement
• Indications for testing:
• Units:g/l or mg/l
• suspected SLE (C3,C4)
• Normal ranges:
• Suspected C deficiency (C3,C4,
haemolytic C)
- C3: 0,68-1,80 g/l
- C4: 0,18-0,60 g/l • Suspected hereditary
angioedema (C2,C4, C1esteraze
inhibitor – immunochemical and
functional)
• Monitoring SLE and immune
complex disease (serum sickness)
 Complement
• Interpretation:
- Complement deficiency is common (especially C2 and
C4 deficiency) predisposes to reccurent neisserial
disease, bacterial infections (C3def.) and immune-
complex disease (lupus-like)
- Measurement of C3 is valuable in monitoring SLE
- ↓ C4 (intercritical), C2 (during acute attacks) – C1-
INH deficiency
• !C3 and C4 are acute-phase proteins (may be normal
even at times of rapid consumption)
C1 component
C1-esterase inhibitor (immunochemical and
functional)
• Units:
• immunochemical – g/l
• functional, reported as percentage activity compared
to normal fresh plasma

• Normal range:
• immunochemical: 0,18-0,54 g/l (pediatric ranges not
well defined, but lower than adult ranges)
• functional: 80-120% normal plasma
Hereditary AngioEdema (HAE)
 C1-esterase inhibitor

• Indications for testing:

• Key indication is angioedema occuring without
urticaria at any age
• Normal C4 during an attack excludes C1-esterase
inhibitor deficiency
• HAE screening test = C4
Rare causes of aquired C1-INH deficiency

• SLE
• Mieloma
• Lymphoma
• Neoplastic disease
Case presentation
 Case history
• 18-years-old woman presents with upper lip swelling for 4 days. The
patient also noted hand swelling for the same duration.
• Medical history
- Patient had had similar symptoms in the past, starting at the age of
7. Over the past 11 years, the episodes had increased in frequency
to the point that at the time of presentation the patient
experienced symptoms three times a month lasting 7 to 10 days per
episode.
• Family history
- The patient’s father, sister, paternal grandfather, paternal
uncle, and two paternal cousins had a history of angioedema.
• Social history - noncontributory. The patient has no known drug
allergies.
 Physical exam

• Normal vital signs

• Periorbital edema, and
upper lip edema
• No other pathological
findings
What is your differential
diagnosis?
 Differential diagnosis

• Drug/food/Insect venom allergy

• Hereditary angioedema
• Aquired angioedema (lymphoma,
mieloproliferative disorders, autoimmunity)
 Skin prick testing (STP)
• The most cost-effective method for
determining whether someone is sensitized to
an allergen
• Principles of testing
• dependent of the release of histamine from
sensitized mast cells.
• Advantages: fast (the patients see the results
as they develop, 15 min to read)
Type I hypersennsitivity
• What Ab do you think is helpful for diagnosis?