J. clin Path. (1967), 20, 826

Incidence and significance of argentaffinand Paneth cells in some tumours

of the large intestineN. M. GIBBS

From Area Laboratory, St. Luke's Hospital, Guildford, and the Research Department, St. Mark's Hospital,London

SYNOPSIS The incidence of argentaffin and Paneth cells in epithelial tumours of the large intestine wasinvestigated. Argentaffin cells were found in adenomatous polyps, villus adenomas, polyposis coli,Peutz-Jehgers' polyps, juvenile polyposis, and adenocarcinomas. Paneth cells were not found inmetaplastic or juvenile polyps.The crypt unit was destroyed in neoplasia and argentaffin and Paneth cells occurred either as a

result of sequestration or were taking part in the neoplastic process.The crypt unit was retained in the disorders of epithelial growth. The identification of argentaffin

and Paneth cells enabled the crypt to be defined and thus provided a useful, practical aid in thedifferentiation between neoplasms and disorders of epithelial growth.

Argentaffin cells are normally found at the base ofthe glands thoughout the entire large intestine.Paneth cells are scanty in the normal large bowelbut may be found in the vicinity of benign andmalignant tumours (Schmidt, 1905; Eklof, 1914;Watson and Roy, 1960). They are also present ingreatly increased numbers in some chronic inflam-matory conditions, particularly ulcerative colitis(Paterson and Watson, 1961).

Feyrter (1931) found argentaffin cells in polyps ofthe large intestine but they were rather scarce.Hamperl (1927) examined 51 carcinomas of theintestinal tract and found argentaffin cells in threecases, but did not specify the site of these tumours.Paneth cells have been described in occasionaladenomatous polyps (Schmidt, 1905; Kerr andLendrum, 1936; Watson and Roy, 1960; Holmes,1965) but Feyrter (1931) found that they were oftenpresent. Paneth cells have been reported only oncein a case of colonic adenocarcinoma (Holmes, 1965).There is no detailed study in the literature of

argentaffin and Paneth cells in tumours of the largeintestine. The object of this research is to investigatethe incidence and significance of both types of cellin a large series of tumours from the large intestinerepresentative of the histological varieties. Morson

Received for publication 20 March 1967.

(1962) discussed the classification of mucosal polypsand for the purposes of this investigation the histo-logical criteria described are followed, particularlyin relation to juvenile, metaplastic, and Peutz-Jehgers' varieties.

MATERLAL

The following tumours were investigated: adenomatouspolyp, papillary adenoma, polyposis coli, adenocarcinoma,metaplastic polyp, Peutz-Jehgers' polyp, juvenile polyp,and juvenile polyposis. The material was obtained fromthe Department of Pathology, St. Mark's Hospital,London, and the Area Laboratory, Guildford andGodalming Group Hospitals. Carcinoid tumours wereexcluded as they have been previously considered (Gibbs,1963).The tissues were fixed immediately in 10% formal

saline. A single block was selected which included thecentre of each tumour, and the tissue was processed andembedded in paraffin wax. Matching sections were cut atSt and stained routinely by Ehrlich's acid haematoxylincounterstained with eosin. Special stains includedphloxine tartrazine, phosphotungstic acid haematoxylin(Mallory) and Gram stain (Weigert's modification) forPaneth cells; Fontana's silver impregnation, Schmorl'sreaction, and the diazo method for enterochromaffingranules; and Perls's reaction for haemosiderin.The following guide to the incidence of these cells in

the tumours was used. The number of glands in each 5 I&section which contained argentaffin and Paneth cells was

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Incidence and significance of argentaffin and Paneth cells in some tumours of the large intestine 827

counted where possible. In addition the total number ofargentaffin and Paneth cells in each section was alsocounted. The average number of argentaffin and Panethcells present in these glands was calculated and expressedas the number of cells per gland section. The resultsobtained from the positive tumours were added togetherand an average figure was calculated for each of thespecial groups of tumour under consideration.

Argentaffin cells were demonstrated by Fontana'ssilver impregnation and the diazo method for entero-chromaffin granules. In general the criteria laid down byLillie and Glenner (1960) were followed to obviatetechnical failure in the argentaffin reaction. Argentaffincells were identified in the normal mucosa adjacent to thetumour in every case.There is no specific stain for Paneth cells and many

staining techniques have been described. In generalphloxine tartrazine, phosphotungstic acid haematoxylin(Mallory) and Gram stain (modified Weigert) provedmost reliable. However, occasionally the staining reactionswere a little inconsistent and variable so that in practicematching sections were stained by each technique andcompared. It was found that the granules of both argenta-ffin and Paneth cells stained a deep red with phloxinetartrazine although the characteristic size and positionof the granules within the cells usually precluded anyconfusion. Phosphotungstic acid haemotoxylin stainedPaneth granules a deep blue and argentaffin granules adiffuse smoky bluish-grey. The Gram (modified Weigert)stained Paneth cells a deep violet colour, while argenta-ifin granules generally were not visible. For the purpose ofthis investigation characteristic staining by each of thesemethods was considered necessary for the positiveidentification of Paneth cells.

ADENOMATOUS POLYP

Argentaffin cells were found in 19 (41 %) of the 46adenomas examined (Fig. 1 and Table). They werelocated in the basal part of the gland when present in

D ARGENTAFFIN CELLS PANEI

100] H H F

w

wz

w

wk

TH CELLS

80

70

60

50

40

30

20

10

I numbers. Sometimes argentaffin cells wereerous and were scattered thoughout the tumour.tumours showed giant argentaffin cells (Fig. 2).th cells were found in nine (19%) of the 46Lomas examined (Fig. 3) and in seven argentaffinwere present. (Fig. 1 and Table).

PAPILLARY OR VILLUS ADENOMA

ntaffin cells were found in nine (39%) of the 23urs examined and they appeared normal.th cells were present in eight tumours and inthey were very numerous (Figs. 1 and 4, ande).

POLYPOSIS COLI

ity-three polyps from 11 cases of polyposis coliexamined and argentaffin and Paneth cellsfound in eight adenomas (Fig. 1 and Table).ntaffin and Paneth cells were not seen in foururs which had undergone malignant transform-. One case was selected for detailed examin-of adenomas from all parts of the large

tine but argentaffin and Paneth cells were founde distributed fairly evenly in all parts of the1 although the incidence was higher in thel polyps which were, however, larger in sizethe proximal polyps.

MALIGNANT TUMOURS OF THE LARGEINTESTINE

hundred and fifty primary adenocarcinomas oflarge bowel were examined. Argentaffin cells6) were found in three tumours (vermiformndix, caecum, and rectum). Paneth cells wered in four tumours (Figs. 7a and 7b). In one case

LI)0 <>. 0 FIG. 1. Theincidence of argentaffinzg ) and Paneth cells in tumours of the

J 0 large intestine.z 0> wD

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N. M. Gibbs

FCi

9~~~~~~~~~~~~~~~~~~~~~. . M

0~~~~~~~~~~~~~~~~~~~2.*~~~~~~~~~~~~~~~~~~~~~FG. 4. Villus adenoma of rectum with Paneth cells.

(Phloxine tartrazine x 80.)

'IG 3. Adenoma of' rectum showing giant argentaffin-ells. (Diazo x .100.)

-AR.-~~~ ~ ~ ~ ~ ~ ~ FG.. Adno:F:A, .~

p~~* * oa frcuO',~~~~~~~~wtPaneth~A

g%t cells (Phioxin-,v:~ ~ ~ )L atrznx 0.

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Incidence and significance of argentaffin and Paneth cells in some tumours of the large intestine

FIG. 5. Polyp from a case ofjuvenile polyposis showingmultiple gland crypts, containing Paneth cells, opening intoa cyst. (Phloxine tartrazine x 63.)

(adenocarcinoma of the caecum) they were scantyand situated near the growing edge, and in theremainder (caecum, transverse colon, and sigmoidcolon) they were found in great numbers (Figs. 7aand 7b). One of these seven tumours (adenocarcin-oma of caecum) contained both argentaffin andPaneth cells. All the positive tumours producedcopious secretion of mucin.

FIG. 6. Adenocarcinoma of rectum with numerousargentafijn cells. (Fontana's silver counterstained withsaffranin x 150.)

METAPLASTIC POLYPS

Thirty-four metaplastic polyps were examined(Fig. 1 and Table) and argentaffin cells were seen inthe gland bases in similar numbers to the adjacentnormal mucosa. Most of the argentaffin cells ap-peared enlarged and the cytoplasm was eosinophilic.Paneth cells were absent.

Paneth Cells

Tumours Cell Count

Adenomatous polyp

Papillary adenoma

Polyposis coli

AdenocarcinomasMetaplastic polypsPeutz-Jehgers' syndromeJuvenile polyps

Juvenile polyposis

46 19 20-6 cells in11-6 gland sections

23 9 8-0 cells in3 0 gland sections

11 8 6-5 cells in23 polyps 3-0 gland sections150 3 Numerous34 34 23-2 cells per tumour9 9 In normal numbers15 15 10-8 cells in

7-7 gland sections3 3 In small numbers

9

8

8

40

30

20-1 cells in4-9 gland sections60-0 cells in13-0 gland sections6-9 cells in2-6 gland sectionsNumerous in 3 tumours

In large numbers

In small numbers butoccasionally numerous

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TABLECases Argentaffin Cells

Tumours Cell Count

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FIG. 7a.

FIG. 7. Adenocarcinoma of sigmoid colon with Panethcells. (a) Ehrlich's haematoxylin and eosin showing num-erous Paneth cells ( x 250); (b) Mallory's phosphotungsticacid haematoxylin ( x 400).

FIG. 7b.

PEUTZ-JEHGERS' SYNDROME

Nine polyps from nine cases of Peutz-Jehgers' syn-

drome were examined (Fig. 1 and Table). Argentaffincells were found in the basal part of most glands andPaneth cells were also present in very large numbersin three polyps.

JUVENILE POLYPS

Fifteen polyps were examined and argentaffin cellswere present in gland crypts in every case. Theargentaffin cells appeared normal in size in spite ofenlarged epithelial cells which formed the tumour.Paneth cells were not found.

JUVENILE POLYPOSIS

Polyps from three cases of juvenile polyposis(McColl, Bussey, Veale, and Morson, 1964) wereexamined. Argentaffin and Paneth cells were presentin small numbers in the majority of polyps fromeach of the cases, and these cells were confined tothe gland crypts (Fig. 5).

Argentaffin cells are present in the gland cryptsthroughout the large intestine and occasional Panethcells are also found, particularly in the caecum. Thegland crypt is the site of normal cell regenerationand in neoplasia the normal basiglandular cells ofthe crypt are replaced by tumour cells. This occursas a primary process if the neoplasm is deriveddirectly from basiglandular cells and as a secondaryprocess if the neoplasm arises elsewhere in the glandwhence the crypt cells are displaced by tumour cells.The adenomatous polyps of polyposis coli were

investigated by Lane and Lev (1963) who concludedthat these neoplasms arose from the crypt cells, andby Cole and McKalen (1963) who found that theyoriginated at the superficial part of the gland. Thisapparent contradiction may be resolved by thehypothesis that neoplasia may commence in bothparts of the gland and that the tumours showingnumerous argentaffin and/or Paneth cells are derivedfrom the basiglandular cells of the crypt. It is notsurprising that tumours arisingfrom thebasiglandularcells of the crypt produce argentaffin and Paneth

DISCUSSION

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cells by differentiation as these cells are present innormal crypts. Neoplastic argentaffin and Panethcells may be expected to be scattered throughoutthe tumour instead of being confined to the crypts.The presence of occasional argentaffin and Panethcells in gland crypts occupied by neoplastic cells isexplained by the occasional survival of normalbasiglandular cells which are 'sequestered' amidstneoplastic cells. Sequestration accounts for thepresence of argentaffin cells in five of the 19 cases(26 %) of adenomatous polyps and eight of the ninecases (89 %) of villous adenomas. A similar explan-ation accounts for the presence of Paneth cells inthree of the nine cases (33 %) of adenomatous polypand one of the eight cases of villus adenoma (12 %).In others, argentaffin and Paneth cells are scatteredthroughout the tumours, sometimes in great num-bers, and it is concluded that these cells are part ofthe neoplastic process. Mitoses in Paneth cells andargentaffin cells are not seen, possibly because thecharacteristic granules are not present in the divi-ding stage. However, neoplastic argentaffin andPaneth cells show loss of orientation and bizarreforms. The majority of the tumours with argentaffinand Paneth cells showed prominent mucin secretionin some parts illustrating their differentiation.

Neoplastic argentaffin cells were present lesscommonly in the villous adenoma than in the adeno-matous polyp, while the reverse was found in thecase of Paneth cells. These findings do not suggestany difference between the malignant potential ofthe two tumours. However, no clear differences werepresent between the solitary adenomatous polyp andthe multiple adenomatous polyps of familialpolyposis.

In contrast to the tumours already described wherethe cytology of the gland crypt is destroyed, theremaining tumours are distinguished by the retentionof the basic gland crypt unit. Argentaffin and Panethcells are located in the crypt and their presenceprovides a useful aid in the identification and theassessment of the gland crypt. The gland crypts in

metaplastic polyp, Peutz-Jehgers' polyp, juvenilepolyp, and juvenile polyposis are easily recognizedby this method even though they may be greatlydistorted. This distortion is minimal in metaplasticpolyp, but very conspicuous in juvenile polyposis(Fig. 5) where it is caused by abnormal and floriddevelopment of the superficial part of the gland.Paneth cells are present in juvenile polyposis butabsent in the solitary juvenile polyp suggesting adifference in the growth processes, even though thepolyps have many similarities. The cytology of thecrypt in some Peutz-Jehgers' polyps and in juvenilepolyposis where Paneth cells are found suggests alocal variation in crypt metabolism. These findingssupport the view that this group of tumours arisesas an anomaly of growth.

I wish to thank Dr. B. C. Morson for his advice andencouragement and the consultant staff of the RoyalSurrey County Hospital and St. Marks' Hospital forpermission to study their cases. For technical assistanceI am grateful to Mrs. R. M. Shrubb, Mr. Lloyd Soodeen,Mr. Norman Mackie, and Mr. L. W. French.The expenses of this investigation were defrayed from

a research grant from the South West MetropolitanRegional Hospital Board, and from a block grant to theResearch Department of St. Mark's Hospital from theBritish Empire Cancer Campaign.

REFERENCES

Cole, J. W., and McKalen, A. (1963). Cancer (Philad.), 16, 998.Eklof, H. (1914). Chondriosomenstudien an den Epithel-und Drusen-

zellen des magen- Darmkanals und den Oesophagus-Drisen-zellen bei Sangetieren. J. F. Bergmann, Wiesbaden. (also Anat.Hefte, 51, 1.)

Feyrter, F. (1931). Beitr. path. Anat., 86, 663.Gibbs, N. M. (1963). J. clin. Path., 16, 206.Hamperl, H. (1927). Virchows Arch. path. Anat., 266, 509.Holmes, E. J. (1965). Cancer (Philad.), 18, 1416.Kerr, A. B., and Lendrum, A. C. (1936). Brit. J. Surg., 23, 615.Lane, N., and Lev, R. (1963). Cancer (Philad.), 16, 751.Lillie, R. D., and Glenner, G. G. (1960). Amer. J. Path., 36, 623.McColl, I., Bussey, H. J. R., Veale, A. M. O., and Morson, B. C.

(1964). Proc. roy. Soc. Med., 57, 986.Morson, B. C. (1962). Dis. Colon Rect., 5, 337.Paterson, J. C., and Watson, S. H. (1961). Amer. J. Path., 38, 243.Schmidt, J. E. (1905). Arch. mikr. Anat., 36, 12.Watson, A. J., and Roy, A. D. (1960). J. Path. Bact., 80, 309.

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