This document discusses immunohistochemistry (IHC), which combines immunology, histology, and chemistry to identify tissue antigens using antibodies. IHC allows identification of cell types and origins. The document outlines the principles and history of IHC, the IHC process, common markers used to identify tissues like epithelial, mesenchymal and hematopoietic tissues. Examples of IHC use in identifying lung cancer subtypes, gastrointestinal cancers, breast cancer subtypes, and lymphoma subtypes are provided. In summary, the document provides a comprehensive overview of the field of immunohistochemistry.

1. MODERATOR:- Dr SHAQAL QAMAR
PRESENTER:- Dr ABDUL WAHEED.
post graduate
skims srinagar kashmir

2.  Immuno histrochemistry is an umbrella term that encompasses
many methods used to determine tissue constituents (the antigens)
with the employment of specific antibodies That can be visualized
through staining. When used in cell preparation it is called
immunocytochemistry, a term that some authors use for all methods
entrailing the immunological search of cell antigens ,even when this
involves tissue slices.
 Brandtzaeg stated that immunostaining for cell markers represents
a way to “talk with cells”, because it allows not only the histological
origin of the cell to be identified but also indicates its function in
vivo, when duly investigated with the correct antibodies.

3.  A SYNERGY OF 3 SCIENTIFIC DISCIPLINES
 IMMUNOLOGY
 HISTOLOGY
 CHEMISTRY

4.  Histochemistry is a science that combines the
techniques of biochemistry and histology in the study of
the chemical constitution of tissues and cells.
 Immunology is a science that deals with the
immune system, cell-mediated and humoral
aspects of immunity and immune responses.
 Immunohistochemistry (IHC) is the integration of
the above mentioned disciplines.

6.  The principle has existed since the 1930s
 In I941: Coons et all. have demonsrated antigens on tissue sections
by using an antibody which was linked to a flouresans label.

 1966: Nakane and Pierce as well as Avrameas and Uriel reported
the use of secondary antibody which was conjugated with
peroxidase enzyme.
 1979: Sternberger described the peroxidase-antiperoxidase
method.
 1981: Hsu et all. have defined the method of avidin-biotin-
peroxidase complex which is abbreviated as ABC method...

7.  Using microscopy and immunology together
 Highly specific binding of antibodies to their
own antigens
 Making antibody-antigen complex
microscopically visible

8.  IHC require the availability of biopsies.
 These are processed into sections with a microtome and then the
sections are incubated with an appropriate antibody.
 The site of antibody binding is visualized under an ordinary or
fluorescent microscope by a marker such as a fluorescent dye,
enzymes or a radioactive element.
 This directly linked to the primary antibody or to an appropriate
secondary antibody.

9.  1. Fixing and embedding the tissue
 2. Cutting and mounting the section
 3. Deparaffinizing and rehydrating the section
 4. Antigen retrieval
 5. Immunohistochemical staining
 6. Counterstaining (if desired)
 7. Dehydrating and stabilizing with mounting medium
 8. Viewing the staining under the microscope

10.  TO DETECT:
 Proteins, carbohydrates, nucleic acids, lipids
 Types of the secreting cells
 Membrane antigens
 Structural antigens within the cytoplasm
 Antigen localised in the nucleus
 Immunohistochemistry is frequently used both in
experimental and diagnostical studies
• Light, flourescens, confocal laser scanning or electron
microscope is used in order to visualize the labeled
antibody-antigen complex.

11. Tumors of uncertain histogenesis
Prognostic markers in cancer
Prediction of response to therapy
Unknown primary
Infection study

12.  IHC methods have brought about a revolution in approach to
diagnosis of tumours of uncertain origin.
 A variety of antibodies is chosen to resolve such diagnostic problem
cases.
 The selection of antibodies being made is based on clinical history
and morphological features.
 Immunohistochemistry stain for intermedaite filament are expressed
by tumour cells ( keratin, desmin, vimentin, neurofilament, and glail
fibrillary acidic protiens).

13.  Predicts the prognosis of tumours by identification of
enzymes, tumour specific antigens, oncogens tumour
suppressor genes and tumour cell proliferation markers.
 Allows for clinical staging and histologic grading.
 Identifies the cell type and origin of a metastasis to find
the site of the primary tumour.

14.  Poorly differentiated tumors and mixed carcinomas
 Undifferentiated tumors of unknown origin
 Treatment based on sub-type of cancer: Personalized
Medicine eg., Breast Ca
 Monitoring progress of cancer (Predictive and Prognosis)
 In Malignant Lymphoma,
 In identifying Carcinoid (Neuroendocrine) tumors
 In Cytologic specimens

15. IMMUNOMARKERS CAN BE BROADLY
CLASSIFIED
 INTERMEDIATE FILAMENT MARKERS
 EPITHELIAL RELATED MARKERS
 GERM CELL TUMOUR MARKERS
 NEUROENDOCRINE MARKERS
 MESENCHYMAL TISSUE MARKERS
 BASEMENT MEMBRANE COMPONENTS
 HARMONE RECEPTORS
 ONCOPROTEIN
 CELL PROLIFERATION MARKERS
 LEUKOCYTE ANTIGENS

16.  .CYTOKERATINS-
20 CK known;- type 1- acidic keratin 12 types, CK9 to CK 20
type 2- basic keratin-8 type- ck1 to ck8
 Also divided into- HMWCK (squamous), LMWCK(simple/non squamous)
intermediate MWCK(basal keratins)
 EMA
 CEA {GLANDULAR EPITHELIA}
 P63 {squamous and urothelial}
 BerEP4 { to diffrentiate lung adenoca from mesothelioma}

17.  Skeletal muscle differentiation
 True smooth muscle differentiation
 Partial smooth muscle differentiation
1.desmin {asst with both skeltal and smooth muscle}
2.Actin{divided into muscle and non muscle)
3.myoglobin(O2 binding heme protien, skeltal and cardiac muscle.)
 Identifying smooth muscle differentiate- myogenin and myoD1
 identifying skeltal muscle differentiation-desmin and smooth muscle actin
 Identifying myofibroblasts desmin –ve, SMA tram track +vity

18.  S-100 PROTIEN- (2 sub units alpha beta,Beta-beta –langerhans and schwann
cell)
 CLAUDIN-1:- ( positive in perinuaral cells,perinuromas)
 GLUT-1 ( perineural cell )
 CD57- ( in nk cells t cells ,oligodendroglial cells and schwann cell)
 MARKERS OF MELNOCYTIC DIFFERENTIATION
 HMB-45 ( +ve in melanoma, Pecoma,-tive in NEVI,RESTING MELANOCYTE).
 Melan –A (product of MART-1 GENE)
(Positive in nevi, and resting melanocytes, desmoplastic melanoma)
 MiFT (micropthalmia transcription factor).
 TYROSINASE ( enzyme involved in synthesis of melanin)
 S-100 (highly sensitive for malenoma)
 RECENT MARKER
SOX10,PNL2,MUM1

19.  CD99 (product of MIC2 gene,transmembrane gp)
 CD56 (neurons, astrocytes,glia,nkcells)
 NB-84 (nuroblastoma)
 SYNAPTOPHYSIN and CHROMOGRANIN A
 NSE (neuron specific enolase)
 CK
MARKERS OF VASCULAR DIFFERNTIATION
 CD31
 CD34
 FACTOR –V111
 ULEX EUROPAEUS I
 CD141
 Fli-1
 ERG(prostate ca)
 D2-40 (podoplanin) ( noval marker for lymphotic endothelial cells)

20. MARKERS TO DEFINE
NSCLC

21.  Bhattacharjee A et al. PNAS,2001
 Garber Meet al.PNAS,2001
 Beer DJ et al. nature medicine,2002
Molecular classification in accordance to histology EGFR
mutation 15% ,75 % with BAC component ,gene expression related to cell
growth ,differentiation, cellular survival.

22.  EGFR MUTATIONS
ACTIVATING MUTATION
10% OF Caucasians ,50% asian
Exon 19 in frame deletion (45%)
L858R missence mutation 45%
Less frequent mutation( 10%) in exons 18 to 21.
KRAS MUTATION:-
25% of adenocarcinomas in a North American
population have KRAS, Patients with KRAS mutations appear to have a
shorter survival than patients with wild-type KRAS.
 ALK PATHWAY-Anaplastic lymphoma kinase (chr2q)
Translocation in 3-7% of ADC

23. OTHER MUTATION
HER2:- overexpression in 17-42% of ADC
amplification and mutation in 5 – 10% of ADC
inframe insersion in exon 20
associated to asian /women/non smoking/ADC.
HER2 mutation associated to resistance to EGFR TKI but sensitive
to her2 therapies.
HER4:- Mutation have been reported in 2-3% of ADC
PIK3CA-AKT-Mt or pathway
 overexpression of akt1
MET:-high expression associated to poor survival
3% with amplification.
Involved in 10-20% of resistance to EGFR TK1.
LKB1:- TUMOR SUPPERSSOR gene
Repress mTOR
Collaboration with kras in oncogenesis

24. Immunohistochemistry stains in
squamous cell carcinoma and
adenocarcinoma of lung. H&E:
hematoxylin and eosin; CK:
cytokeratin; TTF-1: thyroid
transcription factor 1. Squamous
carcinomas are typically positive
for CK5/6 and P63, and
negative for CK7 and TTF-1,
with the reverse profile for
adenocarcinoma although this
case of squamous cell
carcinoma demonstrates focal
weak staining for CK7.
SqCC AdCa

25. Napsin A, an aspartic acid protease whose
expression in the lung is regulated by TTF-1, has
also shown promise in helping to differentiate
primary lung from metastatic adenocarcinomas.
While Napsin A expression may also be seen in
normal kidney and in a proportion of renal tumors,
positivity for both TTF-1 and napsin A is a strong
indication that an adenocarcinoma originated from
lung .

26.  Mandatory
EGFR MUTATION (not all are equivalent)
 STRONGLY RECOMMENDED
ALK
KRAS (BRAF)
 TTF1+ → NSCLC, favour ADC
 p40+ → NSCLC, favour SCC

28.  hematoxylin and eosin stain
 pancytokeratin such as AE1/AE3
 less common ck7 and ck20
 NE marker include cd56,chromogranin and synaptophysin
 ki-67 (high proliferative rate)
 TTF-1 positive for extrapulmonary small cell ca

30.  CDH17 immunoreactivity is most commonly seen in colorectal
adenocarcinomas (up to 96%) and a significant portion of gastric,
pancreatic, and biliary adenocarcinomas (25%–50%). It is rarely found in
adenocarcinomas from outside of GI tract (1%–10%)
 SATB2 gene is associated with cleft palate syndrome in humans. However,
the role of SATB2 in the GI tract is still elusive. Recently, Magnusson et al
found that SATB2 immunoreactivity was restricted to the glandular lining
cells of the human lower GI tract, including appendix, colon, and rectum,
and a subset of neuronal cells in the cerebral cortex and hippocampus.
 Compared with CDH17 and CDX2, SATB2 immunoreactivity is much more
selective for CRC and is rarely seen in carcinomas of the esophagus and
stomach

31.  SATB2 AND CDH17 in diagnosis of medullary carcinoma of the colon and GI
NET.
 The overall HER2-positive rate in diffuse-type gastric or GEJ adenocarcinoma
is lower (5%) compared with intestinal type (up to 30%)
 BRAF V600E mutation in CRCs can virtually exclude Lynch syndrome, and
mutation positive tumors are resistant to anti–epithelial growth factor receptor
therapy.(5% to 9% , cetuximab to front-lineTherapy)
 All patients with metastatic colorectal cancer should have tumor tissue
genotyped for RAS mutations (KRAS and NRAS).
 GIST diagnosis, including immunohistochemistry (95% express CD117 and
80% express CD34, ) and molecular genetic testing (for mutations in kit ).(90%
of patients benefit ,imatinib when their tumors have a exon 11 mutation; approximately 50% of
patients ,when their tumors harbor a exon 9 mutation )
 SUCCINATE DEHYDROGENASE–DEFICIENT GIST


32.  Arginase 1 (ARG1)
hepatocyte paraffin 1 (HepPar1) and glypican 3 (GPC3),
,CD10, CD34.
 HCC VARIANT WITH UNUSUAL IHC STAINING PROFILE
Scirrhous HCC is a rare morphologic variant, composed of less than 5% of
HCCs.
Most of the tumors are negative for HepPar1 and positive for
cytokeratin 7 (CK7), CK19, and epithelial cell adhesion molecule
(EPCAM).
 Hepatoblastoma GPC3 and b-catenin.
 HEPATOCELLULAR ADENOMA
hepatocyte nuclear factor 1 alpha (HNF1a)
b-catenin

33. BREAST

34. ER/PR:-
About 75% of all breast cancers are “ER positive.” They grow in response to the hormone
estrogen. About 65% of these are also “PR positive.” They grow into another hormone,
progesterone.
If your breast cancer’s cells have a significant number of receptors for either estrogen or
progesterone, your cancer is considered hormone-receptor positive and likely to respond to
endocrine therapies
Breast cancer tumors that are ER/PR-positive are 60% likely to respond to endocrine therapy.
Tumors that are ER/PR negative are only 5% to 10% likely to respond to endocrine therapy
Her2:-
In about 20% to 25% of breast cancers, the cancer cells make too much of a protein known as
HER2/neu. These breast cancers tend to be much more aggressive and fast-growing.
For women with HER2-positive breast cancers, the drug Herceptin has been shown to
dramatically reduce the risk of recurrence
TRIPLE NEGATIVE:-
Some breast cancers -- estimates range between 10% and 17% -- are known as “triple
negative” because they lack estrogen and progesterone receptors and do not overexpress the
HER2 protein. The majority of breast cancers associated with the breast cancer gene known
as BRCA1 are triple negative.
These cancers generally respond well to adjuvant chemotherapy

37.  Nodular lymphocyte
predominant
CD45+,EMA+ CD79a+
Lymphocyte rich CHL:-
Nodular Sclerosing CHL
MC CHL, LCHL.
CD15+, CD30+,ALK-

38. IN NON-HODGKINS LYMPHOMA
Generic T-cell Vs B-Cell : CD3, CD20, CD45
Folicular Lymphoma Vs Hyperplasia: Bcl2, Bcl6, CD3,
CD10,CD20
Low Grade B Lymphoma: CD3, CD5, CD10, CD20, CD23,
CD43, Bcl2, Bcl6,
MALT Lymphoma: CD3, Cd5, CD20, Bcl2, ISH Kappa and
Lambda
Myeloma: CD138, ISH Kappa and Lambda
Burkits lymphoma:-CD10,CD19,20,22,BCL6
Mantle cell lymphoma:-CD5,CD20,CD43,CYCLIN D1.

40.  HEAD AND NECK CARCINOMA

41.  Understand the role of IHC in neuropathology to:
 –Make the right diagnosis
 –Provide clinically useful information

42. TUMORS
•Neuroepithelial tumors
–Astrocytomas
–Oligodendrogliomas and mixed gliomas
–Ependymomas
–Choroid plexus tumors
–Glioneuronal tumors
–Neuroblastic tumors
–Pineal parenchymal tumors
–Embryonal tumors
More tumors
•Meningiomas
•Peripheral nerve tumors
•Lymphomas
•Germ cell tumors
•Sellar tumors
•Metastatic tumors

43. Oligodendroglioma
•GFAP, S-100 positive
•1p 19q co-deletion by FISH
Neurocytoma
Synaptophysin positive, most GFAP negative
Clear cell ependymoma
•GFAP positive, focally EMA positive
Clear cell meningioma
•GFAP negative, EMA positive
Oligoastrocytoma
GFAP “brings out”the neoplastic astrocytic component

44. Choroid plexus tumors
•Transthyretin and S-100 positive, but less frequent in choroid
plexus carcinomas
•GFAP positive in 25-55% of papillomas, 20% of carcinomas
•Synaptophysin positive

45.  •Gangliogliomas
•DIG (desmoplastic infantile ganglioglioma)
Need to confirm glial (GFAP) and neuronal differentiation (NF,
synaptophysin)
 Oligodendroglioma-like cells: neu-N positive
 •Papillary or rosetted glioneuronal tumors
Neuropil “islands”positive for neuronal markers
 •Supratentorial PNET
Synaptophysin and GFAP positive

46. Meningiomas
EMA generally positive, but may be weak in:
–Fibroblastic meningiomas
–Atypical meningiomas
– Malignant meningiomas
•CEA positive in secretory meningiomas
Beware of mimics:
–hemangiopericytoma: CD31 and CD34
–Schwannoma: nuclear S-100 positivity
OTHER
Sellar tumors
–Adenomas cytokeratin and synaptophysin +
–Routine panel: prolactin, ACTH, GH, LH, FSH

49. • Express CK7 and CAM 5.2
 Focally positive or negative for CK7.
 Negative for amylase .
 Rare may, express synaptophysin and mucin (MUC) 3.
 usually negative for p63 and CK20.
 also positive anoctamin-1/ANO1, (is a calcium activated
chloride channel protein.)

50.  Adenoid cystic carcinoma expresses both ductal and
myoepithelial/basal cell markers, such as
CK7, CAM 5.2, calponin, SMA, SMMHC, p63, SOX10, and
S100.
 Most adenoid cystic carcinomas showed strong and diffuse
expression of c-KIT
 high-grade
elevated Ki-67 and p53 (poor prognosis)
 Histone H3 lysine 9 trimethylation (H3K9me3) and acetylated
(H3K9ac) ( were independent prognostic factors that significantly
influenced overall survival.)

51.  The tumor expresses
S100, mammaglobin, CK7, CK8, CK18, CK19, 34bE12, EMA,
gross cystic disease fluid protein 15 (GCDFP-15), GATA3,
(STAT5a)( signal transducer and activator of transcription 5a )
MUC1, MUC4,
vimentin .
 negative for (AR),(ER), (PR), and (HER2/neu).
 The MIB-1 indices range between 5% and 28%.

52.  Mucoepidermoid Carcinoma
It is usually positive for
 CK5, CK6, CK7, CK8, CK14, CK18, CK19, EMA, CEA, and p63
 is negative for CK20, SMA, muscle specific actin (MSA), and S100
 However focal expression of S100, c-KIT, glial fibrillary acidic protein (GFAP),
and vimentin can be seen in some tumors
 Sams et al20 , p63 Strongly positive for mucoepidermoid carcinomas.
 Myoepithelial carcinomas
vimentin (100%), calponin (75%–100%),
S100 (82%–100%), CK AE1/3 (90%100%),
34bE12 (92%), CAM 5.2 (89%),
pancytokeratin (74%) EMA (21%–100%);
 less frequently express
 caldesmon (50%), SMA(35%–50%),
 MSA (31%), GAFP (31%–50%), SMMHC(30%),
p63 (28%),
 EMA (27%), and Ki-67

53. The tumor frequently expresses
CK AE1/3, CAM 5.2, 34bE12, EMA, p53, p63, vimentin,
bcl2, S100, and
infrequently expresses SMA and GFAP.
CEA(54%)
Salivary duct carcinoma
AR ( more in men than female), GCDFP-15,
CK AE1/3, CK7, 34bE12, CEA, and EMA
( Occasionally,) positive for ER, PR, S100.
Ki-67 expression markedly increases
GATA3, a new marker for breast carcinoma, was detected in all salivary duct
carcinomas.
More than 80% of salivary duct carcinomas show HER2/neu and p53
overexpression, which was correlated to poor prognosis.

54.  Nuclear Protein in Testis Midline Carcinomas
 Nuclear protein in testis (NUT) midline carcinomas are rare, aggressive
malignant epithelial tumors arising from midline structures, including upper
aerodigestive tract, thymus, mediastinum, lung, and bladder. Approximately
half of these tumors occur in the head and neck regions.
 The NUT midline carcinomas are positive for CK7, p40,p63, and EMA and
are negative or only focally positive for CK20. Approximately one-third of
NUT midline carcinomas are positive for CD34. Rare tumors may show
focal weak immunoreactivity for synaptophysin, chromogranin, and S100.

55. NMC SNUC NPC SCN
EBV - - + -
NUT + - - -
P63 + - Or + + - or+
synaptophysin
chromogranin
- - - +

56. HPV
Human papillomavirus (HPV), predominantly HPV 16,infection has been
identified in a unique subset of patients with head and neck squamous cell
carcinomas especially carcinomas of oropharynx and base of tongue.
p16 overexpression was highly correlated with HPV-positive tumors. Patients
with p16-positive squamous cell carcinomas of oropharynx and oral cavity
showed improved local tumor control, improved disease-free survival, and better
overall survival, compared to patients with p16-negative squamous cell
carcinomas.
p16 immunostain was considered a very sensitive surrogate biomarker for HPV
infection,
The 2011 National Comprehensive Cancer Network guidelines recommended
p16 immunostain alone as a valuable prognostic
 marker for patients with oropharyngeal cancers.

58.  MOST COMMON Ag of LGT
 Tumour supressor protien
 Inactivates cyclin dependent kinases that phosphorylate Rb.
 HPV ONCOgenes E6 and E7 can inactivate pRB and thus lead to p16
overexpression.
 P16 overexpression is a surrogate biomarker of hpv infection
 The intensity and distribution of p16 as well as in nuclear versus cytoplasim
localisation are important
 HPV INDEPENDENT MECHANISIM OF P16 OVERexpression OVARIAN
SERIOUS CARCINOMA.

60.  Primary
 Expreses CK7, GCDFP, AND CEA
 Rare cases express ck20. her2/neu
 But not CDX2 ,S-100, HMB45, ER/PR
secondary
of colorectal origin –ck20, cdx2 and cea; rarely GCDFP
Of urothelial origin –ck20 uroplakin and thrombomodulin
Stromal invasion – ck7, p16 overexpression

61. HIGH GRADE VULVA
INTRAEPTHELIAL NEOPLASIA
MIbI, P16
SIMPLEX(DIFFRENTIATED) VIN P53 GENE
VULVAR MESENCHYMAL TUMOUR VIMENTIN ,ER, PR DESMIN,
ACTIN ,CD34
GRANULAR CELL S100 PROTIEN,Inhibin, calretinin
CD 117 help distinguish vaginal GIST
that mimic smooth muscle tumours

63.  ROLE OF IHC
 distinguish dysplasia from benign mimics
 evaluating cautrized margin
 Grading dysplasia
THE MARKER USED MOSTLY WIDELY ARE MIBI AND P16
 MIB1.
Helps distinguish benign squamous lesion from SIL
Less helpful in distinguishing LSIL from HSILs
 P16
positive LSIL progression rates 36-62.2%
 HPV capsidic protien HPVL1
 Detection rate in L-SIL p16 76.5%, proEx 94.1%, both 100%
 Detection rate in H-SIL proEx 78.6% and p16 alone 1oo%

64.  ENDOCERVICAL AIS EXPRESSES
increased MIBI p16 , monoclonal CEA
 not ER PR ,VIMENTIN OR BCL2
 Tubal metaplasia and endometriosis
show cytoplasmic BCL2
 P16 is either negative or focal and weak.

65.  GENERALY ECA expresses
Monoclonal CEA
P16 diffuse strong nuclear and cytoplasmic pattern
 LOW GRADE ECA
CEA can be positive ,though usually in a weak, luminal pattern
P16 can also be expressed, usually weak patchy but occasionally
strong diffuse pattern
RARELY MIB1 P53

66.  Most will express at one neuroendocrine marker such as
synaptophysin , chromogranin ,or cd56
 Ttf1 can be positive in primary cervical neuroendocrine tumours
 Adenoid cystic carcinoma
P63
Positive for keratin and cd117
 Adeniod basal carcinoma
express keratin p63, p16.

68.  Typically express CK7, CA125, EMA, ER ,PR AND VIMENTIN.
 USUALLY negative for CEA and CK20
 P53 AND P16 ARE only occasionally overexpressed( especially
FIGO grade 1,2)
In significant minorities( 33% figo grade 2,3)
Nuclear beta catenin expression
Loss of PTEN
Loss of DNA MMR PROTIENS( mismatch repair proteins –
MLH1,MHS2 MHS 6,PMS2)

69.  EXPRESS CK7,CA125,EMA AND VIMENTIN
 Usualy negative for CEA and CK20,just like endometriod ca
 Typically overexpress p53 andp 16
 Extremely high proliferative indices with ki-67
 Most cases have low level expression of ER and low or absent PR
 Beta-catenin overexpression ,loss of PTEN and DNA MMR are extremely
rare.
 HER2/NEU expression –poor prognosis

70.  Express CK7,CA125 and vimentin
 Usual negative for CEA AND CK20
 P53 And p16 overexpression is rare
 ER and PR expression is low or absent
 Loss of DNA MMR protien expression can be encountered
 Nuclear expression HNF-Ibeta( hepatocyte nuclear factor) may
emerge as a specific marker
 UNDIFFERENTIATED CARCINOMA
 LOSS OF ER/PR,CK
 RETAIN EMA
 ABERRANT EXPRESSION OF DNA MMR PROTIENS

71. ER/PR P53,P16
overexpression
%of ki67
positivity
Endometriod +++ (except
FIGO grade 3)
+ rare
Serous + +++ Very high
Clear cell - + intermediate

72. ER/PR WT1 CK20 CEA TTF1
Endometrio
d
endometru
m
+++ _ _ - -
Serous
endometriu
m
+ + - - -
Serous
ovary/tube
++ +++ - - -
endocervix - - - +++ -
breast ++ - - + -
colorectal - - +++ +++ -
pulmonary - - - +++ +++

73. PAN TROPHOBLASTIC MARKER
Strongly and diffusely with cytokeratins (AE1/AE3)
Inhibin cd10,ck18,and hydroxyl-d-5-steriod dehydrogenase (HSD3B1)
SYNCYTIOTROPHOBLASTS
Strongly HCG
HPL, PLAP
INTERMEDIATE TROPHOBLASTS
Mel-CAM(CD146), HLA –G, PLAP
IMPLANTATION SITE INTERMEDIATE TROPHOBLASTS
express hpl ,hcg
CHORIONIC TYPE INTERMEDIATE TROPHOBLAST
express p63
CYTOTROPHOBLASTS EXPRESS
beta catenin

74.  PLAP can be positive in most type of malignant germ cell tumour, but
it also can stain epithelial neoplasms
 Oct 4 ,CD117, AND D2-40 are the most useful markers for
dysgeminoma
 Oct-4 and cd30 are the most useful markers for embyonal carcinoma
 AFP And glypican-3 are the most useful marker for yolk sac tumour
 Cytokeratin AE1/AE3 stains many malignant germ cell tumour and the
pattern of staining can help with classification

 EMA IS negative in most malignant germ cell tumour

75.  YOLK SAC
AFP POSITIVE IN 75% of schillar duval bodies
Glypican3 most useful 95% sensitive and specific
Positive for broad spectrum CK (AE1/AE3) but –ve for ck7, EMA
 EMBRYONAL CARCINOMA
Ck (AE1/AE3)POSITIVE
MOST striking oct4 +tive and CD30+ve
PLAP often positive
Rarely AFP and hcg +ve
 CHRIOCARCINOMA
Syncytiotrophoblasts stain with hCG
All trophoblasts +ve for CK,-ve for EMA
 GONADOBLASTOMA
+ for PLAP CD117 And oct4
Sex cord cells stain for vimentin, cytokeratin,and inhibin

76.  TERATOMA
 Little role of mature and imature teratoma
Glial fibrillatory acidic protien(GFAP) stain glial tissue
GFRalpha-1 and MLB1 d/d between immature and mature
teratoma
 Great role of IHC in monodermal teratoma
Ttf1 and thyroglobin+ for thyriod tissue
Chromogranin and synaptophysin +ve for carcinoid
CDX2 d/d from colorectal carciniod

77. CK CK5/6 D2-40 Ber-
EP4
MOC3
1
ER WT1
MESOT
HELIOM
A
+ + + - - - +
SEROU
S CA
- - + + + +

78. DEFINITION
A biopsy proven metastatic cancer in absence of
radio graphically or pathologically detectable
primary tumour after an adequate diagnostic
evaluation.

79.  Adenocarcinoma well to
moderate differentiated
 Squamous cell carcinoma
 Poorly differentiated
carcinoma/poorly
differentiated adeno ca
 Neuroendocrine
 Undifferentiated
malignancy
 60%
 5%
 30%
 2%
 3%

80. PSA
+(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
+
+
+
+
+
+
+
Prostate
Lung
Stomach/Pancreas
Breast
Colon
Colon
Stomach / Pancreas
Breast
Ovary
Pancreas,(Ovary serous)
Stomach / Pancreas
Breast / Stomach / Pancreas
TTF-1
GCDFP15
CDX2 / CK20
ER
CA125
Mesothelin
Lysozyme
+
+
+
(-)
(-)
(-)
CDX2
CK 7
Mesothelin
MC5+ (98%)

81.  20 Subtypes of CK with different molecular weight and different
expressions are seen in various cancers and cell types.
 Monoclonal antibodies to specific CK sub type are used classify tumours
according to site of origins
 Ck7 seen in lung breast, ovary and endometrium.( not seen in lower GI
tract)
 CK20 seen in GI epithelium ,urothelium and merkels cells.
 A pattern of CK20+/CK7- strongly suggests GIT neoplasm.
 A pattern of CK20-/CK7+ SUGGEST cancer of lung, breast ovary,
endometrium and pancreatic biliary tract.

82. CK7 +
CK20 +
CK7 +
CK20 -
CK7 –
CK20 +
CK7 –
CK20 -
Urothelial Ca
Pancreatic Ad Ca
OvarianMucinous
Ca
Ad Ca of Bladder
Gastric Ad Ca
Cholangio Ca
subset
Breast Ca
EndoMetrial Ad Ca
EndoCervical Ad Ca
Ovarian Cerous Ca
Lung Ad Ca
Cholangio Ca
LungSmCC
Mesothelioma
Thyroid Ca
SCC of Cervix
SalivaryGland
tumors
Urothelial Ca subset
Colorectal Ad Ca
Markel Cell ca
Gastric Ad Ca
subset
Prostate Ad Ca
RCC
HCC
Mesothelioma
AdrenoCortical ca
NonSeminoma
LungSmCC
minorSubset
Gastric Ad Ca
subset

86. Pan CK
AE1/AE3
CD45
(LCA)
HMB45
or S100
VIM
Carcinoma Positive Negative Negative Negative
Melanoma Negative Negative Positive Positive
Sarcoma Negative Negative Negative Positive
Lymphoma Negative Positive Negative Negative

87.  Oncofetal protien
 Hormones
 Enzymes
 Tumor associated
antigen
 MISCELLANEOUS
 CEA ; AFP
 HCG ; Calcitonin ; catecholamines
 Prostatic acid phosphatase
 ALP, LDH, NSE
 CA-125 ; CA-15.3 ; CA 19.9
 PSA
 Polyamines
 Nucleosides
 Tissue polypeptide antigen

88.  Technical problem
 Expense
 Expertise
 Cannot replace conventional surgical pathology
 Always be interpreted along with conventional morphological
description

89.  In the recent years, knowledge about cancer IHC has increased
tremendously providing great opportunities for improving the management
of cancer patients by enhancing the efficiency of detection and efficacy of
treatment.
 Recent technological advancement has enabled the examination of many
potential markers and renewed interest in developing new markers.
 A comprehensive understanding of the relevance of each marker will be
very important not only for diagnosing the disease reliably, but also help in
the choice of multiple therapeutic alternatives currently available that is
likely to benefit the patients.
 This review provides a brief account on various markers for diagnosis,
prognosis and therapeutic purposes, which include markers already in
clinical practice as well as various upcoming markers.

90. “ The Diagnostic Power of any
Immunohistochemical Procedure is
no Greater than the wisdom of the
Pathologist interpreting it.”
Dr.Allen M.Gown