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At least 19 recordsLinked to original sources
PubMed1994

Estrogen receptor expression in human pituitary: correlation with immunohistochemistry in normal tissue, and immunohistochemistry and morphology in macroadenomas.

Forty-one human pituitary adenoma specimens were examined for the presence of estrogen receptor (ER) messenger ribonucleic acid and protein using a combination of ribonuclease protection assay, [3H] estradiol ([3H]E2) binding, and ER immunohistochemistry. ER messenger ribonucleic acid prevalence was high in PRL-immunoreactive tumors (2 of 2), moderate in GH/PRL tumors (2 of 5), and low or absent (0 of 4) in GH tumors. In the GH/PRL-immunostaining tumors, the presence of the ER was uniformly associated with elevated serum PRL levels. Among the gonadotropin-immunostaining tumors, 10 of 17 were ER positive; within this group, those with gonadotroph adenoma characteristics were ER positive, whereas those with null cell/oncocytic characteristics were ER negative. Of the tumors that did not immunostain for any known anterior pituitary hormones, 3 of 11 were ER positive. ER immunohistochemistry in 14 tumors revealed a 100% correlation with ribonuclease protection assay results, whereas [3H]E2 binding, determined in 9 tumors, showed an 87% correlation. In summary, it appears that PRL and a specific class of gonadotropin-immunostaining tumors (identifiable by specific characteristics on electron microscope) contain ER, whereas GH-immunostaining tumors are ER negative. ER expression in normal pituitary paralleled that in macroadenomas (GH, 2.3%; PRL, 50%; FSH, 70%; LH, 83%; TSH, 4%; ACTH, 1%). The ER-positive tumors represent a subset whose growth and secretory profiles may be influenced by the gonadal steroidal milieu or by pharmacological agents that affect E2 levels or ER function.

Adenoma↗
PubMed2006

Amplification of the HER2 gene in breast cancers testing 2+ weak positive by HercepTest immunohistochemistry: false-positive or false-negative immunohistochemistry?

BACKGROUND: The majority of cases of breast cancer scoring HER2 weak positive (2+) on immunohistochemistry (IHC) using the HercepTest are not associated with amplification of the HER2/neu gene. AIM: To examine the reproducibility of IHC in cases scoring 2+ subsequently shown to have gene amplification by fluorescence in-situ hybridisation (FISH). METHODS: A retrospective analysis of 153 cases referred for FISH confirmation of a weak positive HercepTest (2+) result was performed. Repeat IHC was undertaken in cases with weak positive (2+) referral IHC and amplification of the HER2 gene by FISH. RESULTS: Amplification of the HER2 gene was confirmed in 29/153 cases (19%) scoring 2+ on IHC. Repeat IHC was carried out on 25 IHC 2+ cases: 7 (28%) scored 2+ on repeat IHC, 18 (72%) scored 3+ and were reclassified as strong positive. A heterogeneous expression pattern was present in 3/17 cases scoring 3+. CONCLUSIONS: The majority of HercepTest 2+ results are not accompanied by gene amplification and represent "false positive" IHC in terms of prognostic or therapeutic relevance. A small proportion of HercepTest 2+ scores represent true 2+ IHC positive cases accompanied by gene amplification: a category probably biologically related to 3+ IHC cases. The remainder of cases of HercepTest 2+ accompanied by gene amplification represent a category of referral IHC 2+ weak positive, FISH amplified, repeat IHC 3+ strong positive, best described as "false negative 2+ IHC". This has implications for selection of cases for FISH analysis where weak positive (2+) IHC score is used as a triage for FISH testing, and for testing strategies in referral laboratories undertaking FISH analysis.

Biomarkers, Tumor↗
PubMed1997

[External evaluation of technical quality of immunohistochemistry. Results of a preliminary multicenter study. Immunohistochemistry Commission of the French Association of Quality Assurance in Pathology and Cytology (AFAQAP-IHC)].

Results of a preliminary study on the evaluation of technical quality in immunohistochemistry are reported. This study has involved 11 centers and has concerned immunodetection of cytokeratins and leucocyte common antigen on paraffin sections. The modalities of slide scoring and the information that can be drawn from the results and from the participants data are discussed.

Evaluation Studies as Topic↗
PubMed1994

Aromatic L-amino acid decarboxylase immunohistochemistry in the suprachiasmatic nucleus of the sheep. Comparison with tyrosine hydroxylase immunohistochemistry.

Using antisera against tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC), we have demonstrated the presence of numerous AADC immunoreactive neurons and a few TH immunoreactive neurons, homogeneously distributed throughout the suprachiasmatic nucleus. Similar results have been described in other species. These observations show that this nucleus is able to synthesize trace amines (such as phenylethylamine or tyramine) in addition to dopamine. It is hypothesized that these trace amines are possibly involved in the integration of day length variation in sheep, a species whose reproduction is closely related to photoperiod.

Animals↗
PubMed2006

Evaluating cytochrome p450 in lesser scaup (Aythya affinis) and tree swallow (Tachycineta bicolor) by monooxygenase activity and immunohistochemistry: possible nonlethal assessment by skin immunohistochemistry.

Six-month-old lesser scaup (Aythya affinis) and nestling tree swallows (Tachycineta bicolor) were injected intraperitoneally with beta-naphthoflavone (BNF) in corn oil or in vehicle alone. Liver samples were taken and stored at -80 degrees C until microsome preparation and monooxygenase assay. Skin samples were placed in buffered formalin for subsequent immunohistochemical (IHC) analysis for cytochrome P4501A (CYP1A). Lesser scaup treated with BNF at 20 or 100 mg/kg body weight showed approximately 6- to 18-fold increases in four monooxygenases (benzyloxyresorufin-O-dealkylase, ethoxyresorufin-O-dealkylase, methoxyresorufin-O-dealkylase, and pentoxyresorufin-O-dealkylase). No IHC response was observed for CYP1A in the skin of vehicle-injected ducks, whereas in the skin from BNF-treated ducks, the positive IHC response was of similar magnitude for both dose levels of BNF. Tree swallows injected with BNF at 100 mg/kg, but not at 20 mg/kg, showed significant increases (approximately fivefold) in hepatic microsomal O-dealkylase activities. Cytochrome P4501A was undetectable by IHC response in skin from corn oil-treated swallows, but positive IHC responses were observed in the skin of one of five swallows at 20 mg/kg and four of five swallows at 100 mg/kg. Although these data do not allow construction of significant dose-response curves, the IHC responses for CYP1A in skin support the possible use of this nonlethal approach for biomonitoring contaminant exposure of birds. In addition, the CYP1A signal observed at the bases of emerging feathers suggest that these might provide less invasive sampling sites for IHC analysis of CYP1A.

Animals↗
PubMed1999

[Recommendations for the immunohistochemistry of the hormonal receptors on paraffin sections in breast cancer. Update 1999. Group for Evaluation of Prognostic Factors using Immunohistochemistry in Breast Cancer (GEFPICS-FNCLCC)].

These recommendations address the immunohistochemical evaluation of estrogen and progesterone receptors in paraffin sections of breast cancers. All the components of the procedure are dealt with: indications, sampling, fixation and processing, section adhesion, antigen retrieval, antibodies, controls, analysis and interpretation of immunostaining, report and quality assurance parameters. The purpose of these guidelines is to serve as a basis for standardization of technic and results and to improve quality control.

Breast Neoplasms↗
PubMed2000

The cost-effectiveness of immunohistochemistry.

OBJECTIVE: To evaluate the cost-effectiveness of immunohistochemistry. DESIGN: Using a theoretical decision analytic model, the cost-effectiveness of immunohistochemistry was evaluated in different scenarios depicting the beneficial use of immunohistochemistry. Data regarding the effectiveness of immunohistochemistry were obtained from the medical literature and costs were obtained from Allegheny General Hospital. SETTINGS: The scenarios depicted both private practice and university patients from which anatomic pathology specimens were obtained. Immunohistochemistry was subsequently performed on these specimens. MAIN OUTCOME MEASURES: Cost, life expectancy, diagnostic certainty, ability to predict prognosis, and cost-effectiveness were evaluated. RESULTS: In all scenarios, immunohistochemistry was cost-effective at very low efficacies. Assuming a per-antibody cost of $50 and use of a 5-antibody panel, immunohistochemistry was cost-effective if it resulted in a 1-year gain of population life expectancy in 1 or more of every 200 cases in which it was applied. Alternatively, if the gain in diagnostic certainty by using immunohistochemistry was 10% and the value placed on a percentage gain in diagnostic certainty was $1000, immunohistochemistry was cost-effective if it produced this increase in certainty in 1 of every 40 tests. If a life-year was valued at $50 000, immunohistochemistry was cost-effective if it resulted in a change in patient prognosis of 0.5 years in 1 of every 100 tests. CONCLUSIONS: Using theoretical modeling, immunohistochemistry is extremely cost-effective. These data have implications in an era of managed care when providers attempt to trim laboratory services. Additional studies are needed to evaluate the actual practice efficacy of immunohistochemistry.

Adult↗
PubMed2005

[Non-functioning pituitary adenomas: clinical features and immunohistochemistry].

UNLABELLED: Clinically non-functioning pituitary adenomas do not produce clinical signs of hormonal hypersecretion. Therefore, signs and symptoms will depend on the mass effect of these adenomas over the central nervous system. Their etiopathogeny is complex and their development is probably influenced by several factors, such as hypothalamic hormones (GHRH), growth factors (FGF), proliferation factors (PCNA, and KI-67), protein P53 and the proto-oncogene c-erb-B2. OBJECTIVE: 1) Determining the clinical features of a population of 117 patients treated for clinically non-functioning pituitary adenoma (age, sex, tumor size, number of surgical procedures, development of hormonal deficiency and hyperprolactinemia). 2) Identifying, after the patients had been clinically characterized, those with clinically non-functioning adenomas with positive immunohistochemistry for hypophyseal hormones (PRL, LH, FSH, GH, TSH and ACTH). 3) Determining if the immunohistochemistry of this population was positive for the cellular proliferation factor Ki-67, protein P53 and protein C-erb-B2 and establishing a correlation with tumor size and tumor invasiveness. This will help in the evaluation of the prognostic value of these proliferation factors. 4) Confronting the results of immunohistochemistry using a standard block with the results of immunohistochemistry using a tissue micro-array. METHOD: Study of the clinical features of 117 patients with clinically non-functioning pituitary adenoma (age, sex, tumor size, number of surgical procedures, development of hormonal deficiency and hyperprolactinemia). Immunohistochemical study (H&E) of 39 patients for hypophyseal hormones, protein P53, protein C-erb-B2, Ki-67 to establish their correlation to tumor growth. The next step was a tissue micro-array of the 39 previously studied cases, using immunohistochemistry for hypophyseal hormones, protein P 53, protein C-erb-B2, Ki-67 to establish their correlation to tumor growth. RESULTS: There was no statistically significant difference between males and females with regards to age, tumor size and number of surgical procedures (p=0.279, p=813, p=139 respectively). There is a statistically significant correlation between the size of the tumor, the number of surgical procedures and hormonal deficiency (p=0.032, p=0.223 respectively). There was no statistically significant correlation between a positive immunohistochemistry for protein P53, protein C-erb-B2, Ki-67 and tumor size (r=0.182, p=0.396; r=-0.181, p=0.397; r=0.272, p=0.199, respectively). The tissue micro-array also did not demonstrate a correlation between positive immunohistochemistry for Ki-67 and C-erb-B2 and tumor size, but it showed a statistically significant correlation between a positive immunohistochemistry for p53 and tumor size (r=-0.696; p=001). CONCLUSION: The biological behavior of the clinically non-functioning adenoma is similar for both sexes. The larger the tumor the greater the number of surgical procedures needed. Hormonal deficiency also becomes more significant as the size of the tumor increases. This paper suggests that a positive immunohistochemistry for p53 is negatively correlated to tumor size, thus demonstrating that it has a predictor value. However, a positive immunohistochemistry for Ki-67 and protein C-erb-B2 does not seem to be a prognostic factor for clinically non-functioning pituitary adenomas, as is the case with other neoplasias.

Adenoma↗
PubMed2003

Quantitative detection of micrometastases in the lymph nodes of gastric cancer patients with real-time RT-PCR: a comparative study with immunohistochemistry.

Histologic examination lacks the sensitivity to detect micrometastases in gastric cancer lymph nodes. In the present study, we applied a real-time RT-PCR approach to the quantitative detection of micrometastases in gastric cancer lymph nodes and compared diagnostic power with routine histology and immunohistochemistry. We studied 392 lymph nodes from 21 gastric cancer patients who underwent curative surgery. Real-time quantitative RT-PCR was performed on a LightCycler instrument using a hybridization probe for carcinoembryonic antigen (CEA) and cytokeratin-20 (CK20) as marker genes. Immunohistochemistry with antibodies to wide-keratin was also performed in the lymph nodes to compare the sensitivity and specificity. Median (average) values of CEA mRNA in lymph nodes in patients with histology(+), immunohistochemistry(+)/histology(-), immunohistochemistry(-)/histology(-) and negative control results were 4600 (16000), 200 (400), 0 (9.8) and 0 (0.6), respectively. There were some false-negative results with simple CEA and CK20 real-time RT-PCR due to the presence of low gene-expressing gastric cancers as revealed by CEA and CK20 immunohistochemistry. CEA in combination with CK20 (duplex) real-time RT-PCR partially covered this weakness. Consequently, all 71 histology(+) lymph nodes were positive for duplex real-time RT-PCR as well as wide-keratin immunohistochemistry. Positivity rates by histology, wide-keratin immunohistochemistry and duplex real-time RT-PCR were 18.0% (71/392), 20.9% (82/392) and 25.8% (101/392), respectively. In 2 of 8 patients with pT1N0, positive lymph nodes were observed by real-time RT-PCR but not by immunohistochemistry. These results indicate that duplex quantitative real-time RT-PCR is the most sensitive method for detecting micrometastases and useful for evaluating the prognostic significance of lymph node micrometastasis in gastric cancer patients.

Carcinoembryonic Antigen↗
PubMed1998

Prognostic significance of progesterone receptor immunohistochemistry in endometrial carcinoma.

OBJECTIVE: The aim of this study was to evaluate the prognostic significance of steroid hormone receptors in endometrial carcinoma using immunohistochemical staining for progesterone receptor (PR) and estrogen receptor (ER). METHODS: We evaluated the correlation between PR/ER immunohistochemistry and age, clinical stage, tumor grade, myometrial tumor invasion, and disease-free survival in a series of 92 cases of endometrioid adenocarcinoma. RESULTS: Fifty (54.4%) endometrial carcinomas were PR-positive and 44 (47.8%) were ER-positive. PR immunohistochemistry of endometrial carcinoma was statistically correlated with the International Federation of Gynecology and Obstetrics (FIGO) stage (I, II vs III, IV, P = 0.001), FIGO grade (G1 vs G2 vs G3, P = 0.007), the depth of myometrial tumor invasion (< or = 1/2 vs > 1/2, P = 0.006), and disease-free survival (living vs dead, P = 0.0025). In contrast, ER immunohistochemistry had significant correlations with the depth of myometrial tumor invasion (P = 0.026) and disease-free survival (P = 0.032). Multivariate analysis of PR/ER immunohistochemistry, stage, grade, and myometrial invasion showed that the PR immunohistochemistry was a significant prognostic factor for survival (P = 0.026). CONCLUSION: PR/ER immunohistochemistry was significantly related to survival and PR immunohistochemistry appeared to be the most reliable means for predicting survival in endometrioid adenocarcinoma of the endometrium, independent of other clinicopathological parameters.

Adult↗
PubMed1998

Cholinergic neurons and terminal fields revealed by immunohistochemistry for the vesicular acetylcholine transporter. I. Central nervous system.

Antibodies directed against the C-terminus of the rat vesicular acetylcholine transporter mark expression of this specifically cholinergic protein in perinuclear regions of the soma and on secretory vesicles concentrated within cholinergic nerve terminals. In the central nervous system, the vesicular acetylcholine transporter terminal fields of the major putative cholinergic pathways in cortex, hippocampus, thalamus, amygdala, olfactory cortex and interpeduncular nucleus were examined and characterized. The existence of an intrinsic cholinergic innervation of cerebral cortex was confirmed by both in situ hybridization histochemistry and immunohistochemistry for the rat vesicular acetylcholine transporter and choline acetyltransferase. Cholinergic interneurons of the olfactory tubercle and Islands of Calleja, and the major intrinsic cholinergic innervation of striatum were fully characterized at the light microscopic level with vesicular acetylcholine transporter immunohistochemistry. Cholinergic staining was much more extensive for the vesicular acetylcholine transporter than for choline acetyltransferase in all these regions, due to visualization of cholinergic nerve terminals not easily seen with immunohistochemistry for choline acetyltransferase in paraffin-embedded sections. Cholinergic innervation of the median eminence of the hypothalamus, previously observed with vesicular acetylcholine transporter immunohistochemistry, was confirmed by the presence of vesicular acetylcholine transporter immunoreactivity in extracts of median eminence by western blotting. Cholinergic projections to cerebellum, pineal gland, and to the substantia nigra were documented by vesicular acetylcholine transporter-positive punctate staining in these structures. Additional novel localizations of putative cholinergic terminals to the subependymal zone surrounding the lateral ventricles, and putative cholinergic cell bodies in the sensory mesencephalic trigeminal nucleus, a primary sensory afferent ganglion located in the brainstem, are documented here. The cholinergic phenotype of neurons of the sensory mesencephalic trigeminal nucleus was confirmed by choline acetyltransferase immunohistochemistry. A feature of cholinergic neurons of the central nervous system revealed clearly with vesicular acetylcholine transporter immunohistochemistry in paraffin-embedded sections is the termination of cholinergic neurons on cholinergic cell bodies. These are most prominent on motor neurons of the spinal cord, less prominent but present in some brainstem motor nuclei, and apparently absent from projection neurons of the telencephalon and brainstem, as well as from the preganglionic vesicular acetylcholine transporter-positive sympathetic and parasympathetic neurons visualized in the intermediolateral and intermediomedial columns of the spinal cord. In addition to the large puncta decorating motor neuronal perikarya and dendrites in the ventral horn, vesicular acetylcholine transporter-positive terminal fields are distributed in lamina X surrounding the central canal, where additional small vesicular acetylcholine transporter-positive cell bodies are located, and in the superficial layers of the dorsal horn. Components of the central cholinergic nervous system whose existence has been controversial have been confirmed, and the existence of new components documented, with immunohistochemistry for the vesicular acetylcholine transporter. Quantitative visualization of terminal fields of known cholinergic systems by staining for vesicular acetylcholine transporter will expand the possibilities for documenting changes in synaptic patency accompanying physiological and pathophysiological changes in these systems.

Acetylcholine↗
PubMed2003

The impact of immunohistochemistry on sentinel node biopsy for primary cutaneous malignant melanoma.

Sentinel node biopsy (SNB) has emerged as an accurate means of identifying nodal disease in patients with malignant melanoma. Superselection of pathological nodes has allowed improved pathological staging of disease. The aim of this study was to look at the impact of immunohistochemistry on pathological staging of sentinel nodes. The first 100 patients undergoing SNB for primary cutaneous malignant melanoma were included in this study. Sentinel node harvesting was performed with the aid of preoperative lymphoscintigraphy and the intraoperative use of both a gamma probe and blue dye. If the sentinel nodes contained tumour on either routine pathology or immunohistochemistry, patients were offered a therapeutic lymph node dissection (TLND). Patients underwent no other treatment to the primary lymph node basin if the sentinel node was free of metastases. In all, 95 patients had at least one node identified, and 25 were staged SNB positive and offered subsequent TLND. We found that 76% (19/25) of SNB positive patients were staged positive on routine pathology, and 24% (6/25) were staged with immunohistochemistry. Immunohistochemistry upstaged disease in 8% of patients (6/76). In all, 21 of the patients staged positive with SNB underwent TLND; 50% (8/16) of the patients staged sentinel node positive with routine pathology showed no further disease in the TLND, compared with 100% (5/5) of the patients staged sentinel node positive with immunohistochemistry only (P<0.05). Three patients have developed recurrence within the nodal basin following a negative SNB. The sensitivity of the procedure is currently 89% (25/28), with a mean follow-up of 24 months. Immunohistochemistry is an essential part of identifying micrometastasis in sentinel nodes, upstaging 8% of patients in our series. Patients with micrometastatic disease may well have a different prognosis from those with occult disease, and careful delineation of these patients is required to determine the prognostic influence of micrometastasis.

Humans↗
PubMed1999

Assessment of proliferative activity in breast cancer: MIB-1 immunohistochemistry versus mitotic figure count.

The proliferative activity is one of the most important single prognostic parameters in breast cancer diagnosis and the time-honored measure of proliferative activity, the mitotic figure count, is an integral component of most combined prognostic scores. The detection of the cell cycle-specific antigens Ki-67, and the development of anti-Ki-67 antibodies, including the paraffin-reactive antibody MIB-1, have established immunohistochemical detection of cell cycle-specific antigens as a measure of proliferative activity in breast cancer diagnosis. The current study was performed to correlate mitotic figure counts with the proliferative activity as assessed by MIB-1 immunohistochemistry, taking into consideration the interobserver reliability of 5 pathologists in estimating mitotic figure counts. In 32 consecutive invasive ductal breast carcinomas, mitotic figure counts were performed independently by 5 pathologists. Mitotic activity was expressed as number of mitotic figures per 10 high-power fields and in a 3-tier score according to the Scarff Bloom Richardson system. Immunohistochemistry was performed using MIB-1 antibody, heat-induced epitope retrieval, and the standard avidin-biotin-immunoperoxidase method. MIB-1 immunohistochemistry was assessed in 3 representative 20x fields by semiquantitative estimation (% of tumor cells positive) and by image analysis (number of MIB-1-positive cells/mm2). We found a high degree of interobserver correlation among 4 experienced pathologists, in both mitotic figures per 10 high-power fields and the 3-tier scoring system. We observed significant, albeit weak, correlations between semiquantitative and quantitative MIB-1 immunohistochemistry and the number of mitotic figures per 10 high-power fields (r between .36 and .53), but this significance was lost in 3 of the 5 observers when mitotic activity was expressed in the 3-tier scoring system. This study confirms mitotic figure counting in the hands of experienced pathologists as a valid, reproducible means of assessing proliferative activity in routine breast cancer diagnosis. The statistically significant, albeit only weak, correlation with MIB-1 immunohistochemistry is in agreement with results obtained by others and suggests that MIB-1 immunohistochemistry cannot be translated by a simple conversion factor into combined prognostic scores to replace the time-honored mitotic figure counts.

Antigens, Nuclear↗
PubMed2006

Immunohistochemistry using a Mycobacterium tuberculosis complex specific antibody for improved diagnosis of tuberculous lymphadenitis.

The clinical and histological criteria used to diagnose lymphadenitis caused by Mycobacterium tuberculosis complex organisms have poor specificity. Acid-fast staining and culture has low sensitivity and specificity. We report a novel method for diagnosis of tuberculosis that uses immunohistochemistry to detect the secreted mycobacterial antigen MPT64 on formalin-fixed tissue biopsies. This antigen has not been detected in non-tuberculous mycobacteria. Polymerase chain reaction (PCR) for amplification of IS6110 from DNA obtained from the biopsies was used as a gold standard. Fifty-five cases of granulomatous lymphadenitis with histologically suspected tuberculosis obtained from Norway and Tanzania were evaluated. Four known tuberculosis cases were used as positive controls, and 16 biopsies (12 foreign body granulomas and four other non-granulomatous cases) as negative controls. With immunohistochemistry, 64% (35/55) and with PCR, 60% (33/55) of granulomatous lymphadenitis cases were positive. Using PCR as the gold standard, the classical tuberculosis histology had sensitivity, specificity, positive and negative predictive values of 92, 37, 60, and 81%, respectively, and immunohistochemistry had sensitivity, specificity, positive and negative predictive values of 90, 83, 86, and 88%, respectively. The observed agreement between PCR and immunohistochemistry was 87% (kappa = 0.73). Immunohistochemistry with anti-MPT64 antiserum is a rapid, sensitive, and specific method for establishing an etiological diagnosis of tuberculosis in histologic specimens. Immunohistochemistry has the advantages over PCR of being robust and cheap, and it can easily be used in a routine laboratory.

Adolescent↗
PubMed1998

A comparison of flow cytometry and immunohistochemistry in human colorectal cancers.

In human colorectal cancer it has been reported that some tumours lack the HLA-ABC antigens. This has been interpreted as reflecting tumour escape from the immune system. Earlier data have been obtained by immunohistochemistry. In this study, we compared the expression of HLA-ABC, HLA-DR, CD80 (B7-1) and CD54 (ICAM-1) in 20 tumours using both a conventional immunohistochemistry two-layer technique and multiparameter flow cytometry, gating on an epithelial cell marker. Colorectal cancer tissue used in flow cytometry was dissociated with collagenase, deoxyribonuclease and hyaluronidase. The intensity of expression of HLA-ABC, HLA-DR and CD80 was unaffected by the enzymes, but CD54 was decreased by 30%. The reproducibility of flow cytometry was good. Microscopy of sections revealed that about 5% of each tumour sample consisted of normal epithelium, but even after correction for this, flow cytometry was superior to immunohistochemistry in 33 out of 80 cases, and showed that tumours described as HLA-ABC negative by immunohistochemistry were in fact weakly positive for HLA-ABC. We conclude that flow cytometry and immunohistochemistry are complementary, and that flow cytometry is superior to immunohistochemistry for detecting antigens/epitopes present in low amounts.

Aged↗
PubMed1988

Detection of cytomegalovirus by in situ hybridisation and immunohistochemistry using new monoclonal antibody CCH2: a comparison of methods.

In situ hybridisation, immunohistochemistry, and morphological analysis for the detection of cytomegalovirus (CMV) were compared in routinely processed tissue sections from a patient with acquired immune deficiency syndrome (AIDS) and widespread CMV infection. Both in situ hybridisation and immunohistochemistry with the monoclonal antibody CCH2 labelled all "owl's eye" cells intensely and, in addition, nuclei of some morphologically normal cells. Quantitative evaluation of the results showed that in situ hybridisation and immunohistochemistry with CCH2 were considerably more sensitive than purely morphological analysis, particularly in tissues with only a few cells infected by CMV. It is further shown that immunohistochemistry with CCH2 detected a higher figure of CMV infected cells than in situ hybridisation. In conclusion, both in situ hybridisation and immunohistochemistry are rapid, sensitive, and specific methods for CMV detection. For routine purposes, however, immunohistochemistry seems to be more suitable.

Acquired Immunodeficiency Syndrome↗