Download Cell Surface Blood Group Antigens in Prostatic Carcinoma

Vol. 81 -No. 4
BRIEF SCIENTIFIC REPORTS
15. Ming SC, Goldman H, Freiman DG: Intestinal metaplasia and
histogenesis of carcinoma in human stomach: Light and electron
microscopic study. Cancer 1967; 20:1418-1429
16. Mowry RW: Alcian blue techniques for the histochemical study of
acidic carbohydrates. J Histochem Cytochem 1956; 4:407
17. Naef AP, Savary M, Ozzello L: Columnar-lined lower esophagus:
An acquired lesion with malignant predisposition. Report on
140 cases of Barrett's esophagus with 12 adenocarcinomas. J
Thorac Cardiovasc Surg 1975; 70:826-834
18. Ozzello L, Savary M, Roethlisberger B: Columnar mucosa of the
distal esophagus in patients with gastroesophageal reflux. Pathol
Annu 1977; 12, pt 1:41-86
19. Paull A, Trier JS, Dalton MD, Camp RC, Loeb P, Goyal RK: The
20.
21.
22.
23.
503
histologic spectrum of Barrett's esophagus. N Engl J Med 1976;
295:476-480
Sheahan DG, West AB: Sulfated mucosubstances in Barrett's (columnar cell) esophageal mucosa (abstract). Gastroenterology 1981;
80:1282
Spicer SS: Diamine methods for differentiating mucosubstances histochemically. J Histochem Cytochem 1965; 13:211-234
Thompson JJ, Zinsser KR, Enterline HT: Barrett's metaplasia and
adenocarcinoma of the esophagus and gastroesophageal junction.
Hum Pathol 1983; 14:42-61
Trier JS: Morphology of the epithelium of the distal esophagus in
patients with midesophageal peptic strictures. Gastroenterology
1970; 58:444-461
Cell Surface Blood Group Antigens in Prostatic Carcinoma
PATRICK D. WALKER, M.D., SATISH KARNIK, M.D., JEAN B. DEKERNION, M.D.
AND JAMES C. PRAMBERG, M.D.
Surface blood group antigens are present to some degree in most
epithelia. These antigens frequently are lost during neoplastic
transformation. The authors looked for the presence or absence
of surface blood group antigens in 52 cases of prostatic carcinoma
of various histologic grades using the specific red blood cell
adherence test. The normal prostatic tissue showed a 2+ reaction
in patients with type A or B blood and 0-1+ in type O. The
hyperplastic areas were 4+ for red blood cell adherence in patients with type A or B blood and 0-4+ in type O. In contrast,
all malignant foci were negative for blood group antigens, no
matter what the histologic grade or blood type. (Key words;
Prostate gland; Carcinoma; Surface blood group antigens; Tumor
markers) Am J Clin Pathol 1984; 81: 503-506
PROSTATE CARCINOMA is an important cause of
morbidity and mortality in older men. It is second only
to lung cancer in frequency and is the third leading cause
of neoplastic related deaths among men.4 The annual
mortality rate from prostatic carcinoma is 22.3 per
100,000.4 Because the behavior of prostatic carcinoma
varies widely among individuals, it is difficult to choose
the appropriate therapy and to predict the individual's
prognosis. Surgical staging and histologic grading are
helpful in predicting overall group survival but are not
helpful when applied to the individual patient.
Received June 28, 1983; received revised manuscript and accepted
for publication September 6, 1983.
Presented in part at the Spring Meeting of the American Society of
Clinical Pathologists, March 1981, San Diego, California.
Supported in part by a grant from the Cancer Association of Greater
New Orleans.
Address reprint requests to Dr. Walker: Department of Pathology,
Tulane University School of Medicine, 1430 Tulane Avenue, New Orleans, Louisiana 70112.
Departments of Pathology and Urology, Tulane University
School of Medicine, New Orleans, Louisiana
A method to separate prostatic neoplasms that are histologically similar but prognostically different is needed.
The specific red cell adherence (SRCA) test has been
shown to provide just such differentiation when applied
to other tissues, particularly in urinary bladder neoplasms3
and oral cavity carcinomas.' This test is based on the
fact that many other tissues besides red blood cells contain
the blood group antigens and that a tissue usually loses
its surface blood group antigens at some point during
neoplastic dedifferentiation.5 The SRCA is performed using an immunologic sandwich technic. The tissue is exposed to an antibody directed against the blood group,
and then indicator red blood cells of the same type, A,
B, or O, are applied. Adherence of the red blood cells
reveals the antigen.
The aims of this study were to determine the presence
or absence of blood group antigens in prostatic carcinoma
and to determine if there was a correlation between the
loss of blood group antigens and histologic grade.
Materials and Methods
Fifty-two patients with a tissue diagnosis of carcinoma
of the prostate gland and five patients with histologically
normal prostate glands were selected from the Pathology
files of the Charity Hospital of New Orleans or the Tulane
Medical Center Hospital. There were 22 patients with
well-differentiated adenocarcinoma, 24 with moderately
WALKER ET AL.
504
differentiated adenocarcinoma, and 6 with poorly differentiated adenocarcinoma. Of the patients with adenocarcinoma, 14 had blood type A, 8 had type B, 4 had
type AB, and 25 had type O. Most of the cases of carcinoma also included areas of benign nodular hyperplasia
and foci of normal prostate. Normal prostatic tissue from
younger patients without nodular hyperplasia or neoplasia was obtained from the autopsy files. Neither the
stage nor the grade of the case were known during the
interpretation of the SRCA.
Human anti-A and anti-B sera were obtained from
Ortho Diagnostics Inc. (Raritan, NJ). Ulex europeus extract kindly was provided by Dr. Stuart Bergman (Tulane
University Medical School). Erythrocytes of A, AB, B,
and O systems were obtained from the blood bank, washed
three times in 85% saline, and diluted to produce a 5%
solution.
Specific Red Blood Cell Adherence Test
Slides were deparaffinized and placed in isotonic TRISbuffered saline (pH 7.4) for 15 minutes. The appropriate
antiserum was applied, and the slides were incubated in
a moisture chamber for 15 minutes followed by TRISbuffered saline (TBS) wash, three changes, five minutes
each. Indicator red blood cells were placed on the slides
for 15 minutes and the slides inverted onto two applicator
sticks in a small amount of TBS to allow the uncombined
red blood cells to fall off. The slides were examined microscopically through the thickness of the glass. Controls
included the use of nonisologous antibody and isologous
red blood cells or isologous antibody and nonisologous
red blood cells. In addition, the sections examined had
built-in positive and negative controls. Normal prostatic
epithelium, transitional epithelium, endothelium, and
erythrocytes served as positive controls. Adipose tissue
and smooth muscle served as negative controls.
The slides were graded semiquantitatively on a 4+ system. A 1 + positive result consisted of adherance of occasional red blood cells to greater than 25% of the area
in question. A grade of 2+ and 3+ consisted of adherence
of more red blood cells to a greater and greater area until
a 4+ condition was reached. A 4+ result consisted of
large masses of red blood cells (often obscuring the underlying structures) adherant to 100% of the area under
examination.
A permanent slide method also was used.6 The slides
were treated as in the first technic up to the point of
inversion into TBS. At that time the sections were fixed
in 2% glutaraldehyde for 15 minutes, rinsed in distilled
water, and stained with hematoxylin and eosin.
Results
Malignant foci had consistently negative results in all
cases, no matter what the stage, grade, or blood type (Fig.
A.J.C.P. • April 1984
1). The SRCA varied according to blood type in the benign
areas. The results for blood groups A, B, and AB were
essentially similar. Normal prostatic tissue was 1-2+ and
benign nodular hyperplasia was 4+ (Fig. 2) in the A, B,
and AB blood groups in each case.
The slides from patients with type O blood were somewhat more difficult to interpret. There was a moderate
variability in the normal (0-1+) and the hyperplastic (04+) epithelium. In each slide, there would be large areas
of 4+ staining of hyperplastic glands and 1 + staining of
normal glands. But always there would be patchy areas
in each where the red blood cells failed to adhere. This
is in agreement with most authors in that the Ulex Europus extract staining is somewhat less consistent than
the antisera used for the other blood types. This test would
not be used to discriminate neoplastic from nonneoplastic
areas.
The SRCA results on the normal prostate obtained at
autopsy did not differ from the built-in normal controls.
The permanent slide technic was superior to the inverted slide method in most respects. The former allows
repeat review over time and multiple observer review not
available with the inverted slide method. In addition,
since the slides are stained with hematoxylin and eosin,
the underlying histology is easily visible using the permanent method. The one disadvantage to the permanent
technic is that there is a slight decrease in the intensity
of the red blood cell adherence, which must be compensated for.
Other readings obtained from specific areas on the tissue
sections with all blood types are as follows: transitional
cell epithelium (2-3+), squamous metaplasia (2-3+),
vessel endothelium (4+), and red blood cells (4+).
Discussion
The results of the SRCA test vary from tissue to tissue
and neoplasm to neoplasm. In the oral cavity, benign
epithelium is strongly reactive, carcinoma in situ is variably reactive, and invasive epidermoid carcinoma is negative for SRCA.1 Grade I papillary carcinomas of the
urinary bladder have a mixed response to SRCA. In histologically indistinguishable neoplasms, the tumors with
loss of SRCA show a highly increased frequency of neoplastic recurrence.3,7 There has only been one previous
study of prostate carcinoma and SRCA.2 This work did
not include the histologic grade and was limited to 15
cases, all negative for blood group antigen in the neoplastic
foci.
We have shown that in the prostate, neoplastic transformation is accompanied by complete loss of SRCA and
that the histologic grade of the neoplasm is not related
to the presence or absence of SRCA. Patients with type
O blood are somewhat more difficult to interpret in that
the normal and the hyperplastic areas may be focally
negative. However, the areas of malignancy were con-
•*>.^
*
* *- m.
*%*
i f r J - *Hi~
FIG. 1. Photomicrographs of the prostatic adenocarcinoma and nodular hyperplasia. A (upper, left). Well-differentiated adenocarcinoma is not
stained with the indicator red blood cells. Hyperplastic areas show 4+ staining SRCA technic (XI25). B (upper, right). Moderately differentiated
adenocarcinoma negative for indicator red blood cells with 4+ staining of benign areas. Note the ease of interpretation of the permanent technic
SRCA and hematoxylin and eosin (X250).
FlG. 2. Photomicrographs of nodular hyperplasia. A (lower, left). The hyperplastic areas stain 4+ with the indicator red blood cells. Notice how
difficult it is to even recognize the tissue with this method SRCA technic (XI25). B (lower, right). The hyperplastic areas, showing 4+ staining with
the indicator red blood cells, are much easier to recognize with the permanent technic SRCA and hematoxylin and eosin (X250).
506
WALKER ET AL.
sistently negative in all areas and all patients. The prostate
carcinoma, therefore, is more like oral cavity epidermoid
carcinoma than papillary transitional cell carcinoma of
the urinary bladder in regard to its loss of blood group
antigens.
These observations suggest another possible use for the
SRCA technic in prostatic pathology. The prostatic lesions
that are borderline between hyperplasia and carcinoma
present a difficult clinical problem. It is essentially impossible to predict which patients subsequently will develop an invasive prostatic adenocarcinoma and which
ones will not. It is possible that the SRCA test might help
to differentiate this group of lesions, so-called atypical
hyperplasia, in much the same way that it separates the
borderline urinary bladder and oral cavity lesions. In patients with borderline oral cavity lesions or grade I transitional cell carcinoma of the urinary bladder, a negative
SRCA test predicts an increased risk of recurrence. We
currently are examining cases of atypical prostatic hyperplasia using the SRCA technic in order to determine
A.J.C.P. • April 1984
if this test can predict patients who will develop a malignant neoplasm.
References
1. George DI, Burzynski NJ, Miller RL: Reactive properties of oral
lesions to the specific red cell adherence test. Oral Surg 1979;
47:51-57
2. Gupta RK, Schuster, R, Christian WD: Loss of isoantigens A, B
and H in Prostate. Am J Pathol 1973; 70:439-448
3 Lange PH, Limas C, Fraley EE: Tissue blood group antigens and
prognosis in low stage transitional cell carcinoma of the bladder.
J Urol 1978; 19:52-55
Silverberg E: Cancer statistics, 1983. CA 1983; 33:9-25
Weinstein RS, Coon J, Alroy J, Davidson I: Tissue associated blood
group antigens in human tumors, Diagnostic immunohistochemistry. Edited by RA DeLellis. New York, Masson Publishing,
1981, pp 239-261
Yamase HT, Powell GT, Koss LG: A simplified method of preparing
permanent tissue sections for the erythocyte adherence test. Am
J Clin Pathol 1981;75:178-181
Young AK, Hammond E, Middleton AW: The prognostic value of
cell surface antigens in low grade, non-invasive transitional cell
carcinoma of the bladder. J Urol 1979; 122:462-464
Electronic Counting of Spinal Fluid Cells
INGEBRIGT TALSTAD, M.D.
An analysis of the electronic counting of leukocytes in cerebrospinal fluid (CSFLpc) was made theoretically in models and in
patients. At spinal fluid dilutions of 1/500, 1/50, 1/25, and
1/2, linearity was obtained by electronic counting down to 1,000,
100, 20, and 2 Lpc (106/L), respectively. The electronic particle
counters produce satisfactory results at Lpc > 100 (106/L) but
need modifications to produce satisfactory results at Lpc < 100
(106/L). It is theoretically possible to reduce the variation nine
times by electronic counting as compared with microscopic
counting. A method for the correction of blood admixture at
traumatic spinal puncture by electronic counting was shown to
be satisfactory. (Key words: Spinal fluid; Electronic counting)
Am J Clin Pathol 1984; 81: 506-511
MICROSCOPY is at present the routine method for spinal
fluid cell counting, however, the precision is poor at low
cell numbers. The precision is improved by ultrafiltration
of a large volume of spinal fluid,1 but this method is
cumbersome in routine use. While the electronic counting
Received March 25, 1983; received revised manuscript and accepted
for publication October 24, 1983.
Address reprint requests to Dr. Talstad: 5016 Haukeland Hospital,
Bergen, Norway.
Hematological
Division, Haukeland Hospital,
University of Bergen, Norway
of blood cells is a routine method, no similar method is
yet available for spinal fluid cell counting. The present
study demonstrates a theoretic approach to spinal fluid
cell counting and studies the capacity of electronic particle
counters to measure the low cell concentrations that may
occur in spinal fluid.
Materials and Methods
Theoretical Approach
The standard deviation (SD) in microscopic or electronic counting equals the square root of the number of
cells counted (Poisson's law).2 The relative precision of
electronic counting theoretically is improved compared
with microscopic counting by using a lower dilution of
the spinal fluid and/or by counting a larger volume
(Table 1).