Fabre, Michel

Grosshans, Edouard

Hanau, Daniel

Stampf, Jean-Luc

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LANGERHANS CELLS IN MOUSE EPIDERMIS 

290 LETTERS TO THE EDITOR 
TK: Determination of trimethylpsoralen in blood, ophthalmic 
fluids, and skin. J Invest Dermatol 79:347- 377, 1982 
3. Chakrabart i SG, Pradhan TK, Grimes PE, Minus HR, Diolulu M, 
Kenney JA Jr: Distribution of 4,5' ,8-trimethylpsoralen and me-
tabolites in body fluids and tissues (abstr). J Invest Dermatol 
80:307, 1983 
4. Murata T, Toshioka N, Kito H: Studies on absorption, excretion, 
distribut ion and metabolism of 3H-trioxsalen. P harmacometrics 
4:577- 582, 1970 
5. Mandula BB, Pathak MA, Dudek G: P hotochemotherapy: identi-
fication of a metabolite of 4,5' ,8-trimethylpsoralen. Science 
193:1131- 1134, 1976 
6. Mandula BB, Pathak MA: Metabolic reactions in vitro ofpsoralens 
with live r and epidermis. Biochem Pharmacol 28:127- 132, 1979 
7. Kaidbey KH, Kligman AM: Photopigmentation with t rioxsalen. 
Arch Dermatol 109:674-677, 1974 
8. Sehgal VN: Oral trimethylpsoralen in vitiligo in children: a pre-
liminary report. Br J Dermatol 85:454- 456, 1971 
9. Sehgal VN: Trioxsalen t herapy for vitiligo. Arch Dermatol 109:578, 
1974 
10. Sehgal VN: Effectiveness of t rioxsalen therapy for vitiligo. Arch 
Dermatol 110:957- 958, 1974 
Siba G. Chakrabarti , Ph.D. 
Pearl E. Grimes, M.D. 
Harold R. Minus, M.D. 
John A. Kenney, Jr. , M.D. 
Tapas K. Pradhan, Ph.D. 
Department of Dermatology 
Howard Un iversity College of Medicine 
Washington, D. C. 
LANGERHANS CELLS IN MOUSE EPIDERMIS 
To the Editor: 
We have read with great interest the article by K. W. Baker and J. 
E . J. Habowsky on "EDTA Separation and ATPase Langerhans Cell 
Staining in t he Mouse Epidermis" (J Invest Dermatol 80:104- 107, 
1983). In thei r study the authors point out t hat the technique used by 
Mackenzie and Squier [1] ensures a good definition of cell bodies and 
dendrites of Langerhan s cells (LC). Their Fig 4, however, shows a 
FIG 1. Epidermal sheets stained for ATPase activi ty. Many- Lan 
gerhans cells are visible with thin and ramified dendrites (X 160). 
shrunken aspect _of LC cell bodies. Moreover, under the electron micro-
scope- though with a different technique from the one recommended 
by Mackenzie and Squier-they remarked t hey "have been unable to 
observe intracellular Langerhans granules ... an observation which 
suggests that Trismal buffer, at pH 7.3, is relatively unsui table for t he 
optimum preservation of epidermal ultrastructure ... . " 
May we report our own experience on t he same subject. We used, 
although with t he guinea pig, the method of Mackenzie and Squier for 
t he study of LC with a 5-min incubation period in ATP, as t hese 
aut hors suggest. 
Vol. 81, No.3 
The images obtained under the light microscope a re very fine (Fig 
1): t he cells are clearly marked, unshrunken, and many of the dendrites 
are t hin and ramified. We continued our study using electron micros-
copy, still fo llowing Mackenzie a nd Squier (method of Hirsch and 
Fedorko), and found a regular though discontinuous labeling (lead 
sulfide) around a number of cells, with a clearer cytoplasm than that 
of the neighboring keratinocyte. In t he cytoplasm of these clearer cells 
we could observe no Langerhans granules, but scattered unorganized 
membranous structures. We then proceeded to t he following modifi-
cations: (1) adjunction of a 5% calcium chloride solut ion (0.2 ml per 
100 ml) in the Trismal buffer and t he cacodylate-buffe red fo rmalde-
hyde, (2) reduction from 20 min to 2 min of the t reatment duration in 
dilute ammonium sulfide at room temperature, (3) fixation of the 
fragments after revelation of the membranous ATPase activity in 
Takahashi medium [2]. 
FIG 2. A dendrite whose limits a re labeled by lead sulfide. It conta ins 
a Langer hans granule (arrow) (X 30,000). Inset, A higher magnification 
of t he Langer hans granule (X 100,000). 
Electron microscopy then showed not on ly the membranous ATPase 
reaction product but also well-preserved intracellular organelles, in 
particular Langer hans granules (Fig 2). Throughout our study, however, 
we used Trismal buffer! We therefore think t hat even though Trismal 
buffer may not be t he most adequate medium, other media, in particular 
the one employed fo r the fixation, can influence the quality of the 
results in electron microscopy. 
REFERENCES 
1. Mackenzie IC, Squier CA: Cytochemical identification of ATPase-
positive Langerhans cells in EDTA-separated sheets of mouse 
epidermis. Br J Dermatol 92:523- 533, 1975 
2. Takahashi G: Simul taneous fixation with glu taraldehyde, picric 
acid, osmium tetroxide mixture for electron microscopy, Pro-
ceedings of the 7th European Congress on E lectron Microscopy, 
Leiden, 1980, vol II. Edited by P Rederoo, W De Priester. pp 
746- 747 
Daniel Hanau, M.D. 
Michel Fabre, M.D. 
Jean-Luc Stampf, P h.D. 
Edouard Grosshans, M.D. 
Universite Louis Pasteur 
Strasbourg, France 
REPLY 
We would like to thank Dr. Hanau, Dr. Fabre, and Dr. Stampf for 
their comments concerning the adenosine triphosphatase (ATPase) 
staining of epidermal sheets. 
As pointed 0\-!t in our recent art icle (J Invest Dermatol 80:104- 107, 
1983), t he epidermis of the mouse differs from that of the guinea pig 
in a number of respects, most significant ly, in t he vertical disposition 


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LANGERHANS CELLS IN MOUSE EPIDERMIS

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