Centre of Excellence for Diagnosis Of Primary Immunodeficiency Disorders (PID)
- Established state-of-the-art facilities for evaluation of suspected cases of Inborn Errors of Immunity (IEIs).
- Evaluated a total of 11,510 suspected Primary Immunodeficiency (PID) cases, with confirmed diagnosis in 909 cases.
- Based on screening results, specialized assays and genetic confirmation using Sanger sequencing or next-generation sequencing (NGS) were performed.
- Established normal reference ranges for rare lymphocyte subsets in the Indian population.
- Set up facilities for prenatal diagnosis (PND) using cordocentesis, amniotic fluid, and CVS samples via flow cytometry and molecular techniques.
- PND offered in 56 families (64 pregnancies): 32 cases with phenotypic diagnosis via cordocentesis and 32 via molecular analysis.
- Prepared standard operating procedures (SOPs) for common diagnostic tests for PIDs.
- Drafted a policy brief to the Government of India recommending free IVIg therapy for patients with hypogammaglobulinemia.
- Organized annual CME-cum-flow cytometry training workshops since 2011 to create awareness and build diagnostic capacity across India.
- Conducted 13 training programmes, training over 800 participants nationwide, along with numerous CMEs and PID awareness programmes in various centers.
Established Diagnostic facilitates for IEIs Lymphocyte subset analysis:
I. T Cells and Their Subsets
- T helper (CD4+)/ T cytotoxic (CD8+)
- Naïve (CCR7+/CD62L+/CD27+/CD45RA+)
- Memory (CCR7+/CD62L+/CD27+/CD45RA-)
- Effector (CCR7-/CD62L-/CD27-/CD45RA-)
- TEMRA (CCR7+/CD62L+/CD27+/CD45RA-)
- TsCM (CCR7+CD45RA+CD28+CD95+)
- CD8+CD28- T cell
- Senescent (CD57+CD28-)
- Exhausted (PD1+/Tim3+)
- T follicular helper (CXCR5+CD45RA-PD1+)
- Th1/ Tc1-like (CXCR3+CCR4-)
- Th2/ Tc2-like (CXCR3-CCR4+CCR6-)
- Th17/ Tc17-like (CCR4+CCR6+CCR10-)
- Th9 (CCR4-CCR6+)
- Th22 (CCR4+CCR10+CCR6+)
- Recent thymic emigrants (CD31+CD62L+CD45RA+)
- αβ+ T cells (GD-)
- γδ+ T cells (GD+)
II. Regulatory T Cells and Their Subsets
- Treg (CD4/CD8) (CD25++CD127-/low)
- Memory Treg (CD95+CCR4+CD45RAlowHLADR+)
- Naïve Treg (CD45RA+CCR4-/lowCD95-)
- V beta repertoire analysis
III. B Cells and Their Subsets
- Naïve (CD27-IgD+)
- Memory (CD27+IgD+)
- Class-switched memory (CD27-IgM-IgD-)
- Germinal centre (CD27+IgD-)
- Marginal zone (CD27+IgD+IgM+)
- IgM-only memory (CD27+IgD-IgM+)
- CD21 low (CD21-/CD38-)
- Plasmablast (CD27+CD24+CD38+)
- Activated B cell (CD95+CD38-)
- B regulatory (CD24++CD38++)
- Transitional 1 (CD27-IgD+ CD24+ CD10+ CD38+)
- Transitional 2 (CD27-IgD+CD10-CD38+)
- Transitional 3 (CD27-IgD+CD10-CD38-)
IV. Double Negative T (DNT) Cells and Their Subsets
- B220+ : Proliferative DNTs
- CD27+CD28+ : Naïve DNTs
- CD45RA-CD27- : Effector DNTs
- CD57+KLRG1- : Senescent DNTs
Established Diagnostic facilitates for IEIs Lymphocyte subset analysis:
Surface Protein Expression for the Diagnosis of Specific IEIs
- Btk: X-linked agammaglobulinemia
- CD127: IL-7 receptor α deficiency
- CD132: X-linked SCID
- CD18/CD11a/CD11b/CD11c: LAD-I
- CD40: AR Hyper IgM syndrome
- CD40L: X-linked Hyper IgM syndrome
- DOCK8: DOCK8 deficiency
- WASp: Wiskott-Aldrich Syndrome
- p47, p22, p67: Chronic Granulomatous Disease
- IL12Rb1: MSMD
- IFNgR1: MSMD
- CTLA4: CTLA4 haploinsufficiency
- LRBA: LRBA deficiency
- Perforin: FHL2
- XIAP: XLP2
- FoxP3: IPEX syndrome
- NKg2D expression: XMEN syndrome
Functional Assays
- pSTAT1, pSTAT3, pSTAT4, pSTAT5
- Granule release assay
- NBT/DHR/FMLP
- Apoptosis assay
- CD154 expression
- CD62L shedding
- RBC ADA levels
- T cell proliferation by PHA, anti-CD3, and CD28
Other Specialized Tests
- TREC
- T cell Vβ repertoire
- Immunoglobulin levels (IgG, IgA, IgM, IgE)
- IgG subclass analysis (IgG2, IgG4)
- Specific antibody responses (anti-tetanus, anti-typhoid)
- Complement system evaluation (CH50, AH50)
