Journal of Cutaneous Pathology 1975: 2: 97-102
Some Ultrastructural Aspects of Lymph-Node Cells and Hepatocytes in Papular Acrodermatitis of Childhood F. GlANOITl AND R. CAl'UrO
Clinic of Dermatology, University of Milan, Milan, Italy Ultrastructural examination of the inguinal lymph-nodes of six patients with papular acrodermatitis of childhood with hepatitis B antigeneniia showed the presence of several Langerbans cells and, in three cases, the presence of some mononucleate cells, the cytoplasm of which contained groups of 20-50 spherical particles, approximately 150 A in diameter and with an electron-dense core. In the hepatocytes of two of these patients we observed spherical particles ranging in size between 150 A and 220 A, having an electron-lucent core and scattered throughout areas limited by a membrane. (Received for publication November 14, 1974)
Papular acrodermatitis (PAC) is an infective disease of childhood slightly contagious and fairly widespread, characterized by: a) a non-itching, non-relapsing erythematopapular dermatitis, loealized on the face, buttocks and limbs, lasting 20-25 days; b) an enlargement of inguinal and axillary lymphnodes, due lo a reactive rcticulohistiocylic lymphadenitis; e) an acute hepatitis, usually anieleric, whieh commonly lasts about two months, but may become a ehronic persistent hepatitis or, more rarely, an aggressive hepatitis with moderate activity. This condition possesses all the characteristic features of a diffuse reactive histioeytosis of viral origin and the constant presenee of the HB antigen might signify that it has the same etiology as serimi hepatitis (Gianotti 1973, 1974). After investigating the tillrastrucUire of the cells present in cutaneous papulae (Bellone et al. 1967) and in 23 liver biopsies (Jean et al. 1967) of 16 children suffering
from PAC, we also examined those present in the lymph-nodes of six new cases. Liver biopsies were also studied in two of these cases. The serum of all the patients was examined by the negative-staining technique. Material and Methods
Bioptic fragments of the inguinal subcutaneous lymph-nodes of six children with PAC in progress and with hepatitis B antigenemia were fixed in 1 % osmium tetroxide in Millonig's buffer (Millonig 1962) for 4 h at 4°C and then embedded in Vestopal, according to the teehnique described by Caputo & Lombardi (1963). The sections obtained with a LKB Ultratome mierotonie and stained with uranyl acetate and lead cilratc were observed with a Philips EM 200 microscope. The same method was used for the liver biopsies of two of these patients. The serum of the six patients was examined with the negative-staining technique.
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Fig. 1. Typical Langerhans cell in a lympb-node of PAC. X 8,500.
Fig. 2. Mononucleate cell of a lymph-node of PAC, containing two clusters of spherical particles. X 43,500.
SOME ULTRASTRUCTURAL ASPECTS OE LYMPH-NODE CELLS
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SOME ULTRA.SIRUCTURAL ASPECTS OF LYMPH-NODE CELLS Results
In addition to numerous mononucleate cells and large maerophagie cells in the six lymphnodes examined, we also observed the presenee of a fairly high number of Langerhans cells (Fig. 1) whieh looked similar to the Langerhans eells normally found in the cutis. The Langerhans granules are generally rod-shaped, rarely tennis raquet-shaped, and never attaehed to the cell membrane. The cytoplasm of these cells usually contains many phagosomes and myelinic figures, lipidic vacuoles very rarely being found, hi some cells the rough endoplasmic reticulum is well developed, while in others the ccntrioles are evident. In the lymph-nodes of three patients, the cytoplasm of mononucleate cells exhibited groups of spherieal particles, approximately 150 A in diameter, whieh eoiisisled of an electron-dense core surrounded by a membrane. These groups ineluded 20-50 elements, sometimes showing an alveolar arrangement (Figs. 2, 3 A). One cell may even contain two or three of these clusters (Fig. 2), some of these partielcs also being observed in intercellular spaces. At no time did we find any Langerhans granules in these cells. In the hepatocytes of two of these patients we observed spherical particles of variable size (between 150 A and 220 A), generally with an electron-lucent core, scattered throughout a space limited by a membrane (Fig. 3B). Pleomorphie spherical particles, 200 A in diameter, tubular forms of the same diameter and of varying length, and double-shelled particles 420 A in diamclcr
101
(Dane particles) were observed in the sera of all six patients (Fig. 3 C).
Discussion Langerhans cells in the lymph-nodes have been so far observed in histioeytosis X (Gianotti et al. 1968), in dermatopathie lymphadenopathy (Jimbow el al. 1969) and in the normal rabbit (Kondo 1969). These eells are always present in the lymph-nodes of PAC, with Langerbans granules usually always rod-shaped and never attached to the cellular membranes. The partiele aggregates found in the cytoplasm of monocytc lymphnode cells are not glycogen clusters, sinee these are totally electron-dense and lack a limiting membrane. Fhc spherical particles we observed in the cytoplasm of hepatocytes differ from those seen in the lymph-nodes in three ways: their size ranges from 150 A to 220 A; their eleetron-lueent core; and they are not elustcred but scattered throughout areas limited by a membrane. From a morphological point of view, these particles are rather similar to those demonstrated by various authors, as reported by Gerber et al. (1974), in the nucleus and in the cytoplasm of hepatocytes of acute and chronic hepatitis. However, in order to be able to prove thai they positively are the image of the HB antigen, a comparative study with immunoclcctron microscopy will be required. The particles evidenced in the blood serum with the negative-staining technique are, on the other hand, typical of the HB antigen. These findings are now conlributiiig lo the
Fig. 3 A. Particles 150 A in diameter with an electron dense core, in a lymph-node of PAC.X 170,000. B. Spherical particles ranging in size between 150 A and 220 A in hepatocytes of PAC. X 135,000. C. The three types of parlicles associated with the hepatitis B antigen, present in the serum of a patient suffering from PAC. X 280,000.
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substantiation of the view that PAC is a clinical disease due to the primary infection of the HB antigen in childhood, acquired via oral or mueous membranes (Gianotti 1974). References Bellone, A. G., Caputo, R. & Gianotti, F. (1967) Aspetti ultrastrutturali cutanei deH'acrodermatite papulosa infantile di Crosti-Gianotti. Minerva Dermatologica 42, 278-279. Caputo, R. & Lombardi, L. (1963) Oniidri ultrastrutturali dell'epidermide umana normale. Atti dclla Accademia medica lombcirda 18, 72-77. Gerber, M., Hadziyamis, S., Vissoulis, C, Scbaffner, F., Paronetto, F. & Popper, H. (1974) Electron microscopy and immunoelectionmicroscopy of cytoplasmic hepatitis B antigen in bepatocytes. American Journal of Pathology 75, 489-496. Gianotti, F., Caputo, R. & Ranzi, T. (1968) Ultrastructural study of giant cells and Langerhans ceils granules in cutaneous lesions and lymph nodes and liver biopsies from four cases of siibacute disseminated hisliocytosis of Letter-Siwe. Arcliiv fiir Klinische und Experimentelle Dermatologie 233, 238-252. Gianotti, F. (1973) Papular acrodermatitis of
childhood. An Australian antigen disease. Archives of Diseases of Childhood 48, 794— 799. Gianotti, F. (1974) Hepatitis B antigen in papular acrodermatitis in children. British Medical Jonrnal 5924, 169-170. Jean, d., Lanibeitengbi, G., Gianotti, F., Travaglini, P. & Ranzi, P. (1967) L'epatite aciita anitterica iiell'Acroderniatite papnlosa infantile. Milan: Ganassini Publ. Jimbow, K., Sato, S. & Kukita, A. (1969) Cells containing Langerhans granules in human lymph nodes of dermatopatbic lymphadenopathy. Journal of Investigative Dermatology 53, 295-299. Kondo, Y. (1969) Macrophages containing Langerhans granules in normal lymph-nodes of the rabbit. Zcitschrift fiir Zellforschnng und Mikroskopische Anatomic 98, 506—511. Millonig, G. (1962) Furlber observations on a phosphate buffer for osmium solution. Proc. 5tli Intern. Congr. of Electron Microscopy, ed. Hieese, S. S., Jr. Philadelphia, p. 8. New York-London: Academic Press. .Address: /';•()/. F. Giatiotti Clinic of Dermatology University of Milan Via Pace 9 Milan, Italy
Some Ultrastructural Aspects of Lymph-Node Cells and Hepatocytes in Papular Acrodermatitis of Childhood F. GlANOITl AND R. CAl'UrO
Clinic of Dermatology, University of Milan, Milan, Italy Ultrastructural examination of the inguinal lymph-nodes of six patients with papular acrodermatitis of childhood with hepatitis B antigeneniia showed the presence of several Langerbans cells and, in three cases, the presence of some mononucleate cells, the cytoplasm of which contained groups of 20-50 spherical particles, approximately 150 A in diameter and with an electron-dense core. In the hepatocytes of two of these patients we observed spherical particles ranging in size between 150 A and 220 A, having an electron-lucent core and scattered throughout areas limited by a membrane. (Received for publication November 14, 1974)
Papular acrodermatitis (PAC) is an infective disease of childhood slightly contagious and fairly widespread, characterized by: a) a non-itching, non-relapsing erythematopapular dermatitis, loealized on the face, buttocks and limbs, lasting 20-25 days; b) an enlargement of inguinal and axillary lymphnodes, due lo a reactive rcticulohistiocylic lymphadenitis; e) an acute hepatitis, usually anieleric, whieh commonly lasts about two months, but may become a ehronic persistent hepatitis or, more rarely, an aggressive hepatitis with moderate activity. This condition possesses all the characteristic features of a diffuse reactive histioeytosis of viral origin and the constant presenee of the HB antigen might signify that it has the same etiology as serimi hepatitis (Gianotti 1973, 1974). After investigating the tillrastrucUire of the cells present in cutaneous papulae (Bellone et al. 1967) and in 23 liver biopsies (Jean et al. 1967) of 16 children suffering
from PAC, we also examined those present in the lymph-nodes of six new cases. Liver biopsies were also studied in two of these cases. The serum of all the patients was examined by the negative-staining technique. Material and Methods
Bioptic fragments of the inguinal subcutaneous lymph-nodes of six children with PAC in progress and with hepatitis B antigenemia were fixed in 1 % osmium tetroxide in Millonig's buffer (Millonig 1962) for 4 h at 4°C and then embedded in Vestopal, according to the teehnique described by Caputo & Lombardi (1963). The sections obtained with a LKB Ultratome mierotonie and stained with uranyl acetate and lead cilratc were observed with a Philips EM 200 microscope. The same method was used for the liver biopsies of two of these patients. The serum of the six patients was examined with the negative-staining technique.
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Fig. 1. Typical Langerhans cell in a lympb-node of PAC. X 8,500.
Fig. 2. Mononucleate cell of a lymph-node of PAC, containing two clusters of spherical particles. X 43,500.
SOME ULTRASTRUCTURAL ASPECTS OE LYMPH-NODE CELLS
99
100
GIANOTTI AND CAPUTO
SOME ULTRA.SIRUCTURAL ASPECTS OF LYMPH-NODE CELLS Results
In addition to numerous mononucleate cells and large maerophagie cells in the six lymphnodes examined, we also observed the presenee of a fairly high number of Langerhans cells (Fig. 1) whieh looked similar to the Langerhans eells normally found in the cutis. The Langerhans granules are generally rod-shaped, rarely tennis raquet-shaped, and never attaehed to the cell membrane. The cytoplasm of these cells usually contains many phagosomes and myelinic figures, lipidic vacuoles very rarely being found, hi some cells the rough endoplasmic reticulum is well developed, while in others the ccntrioles are evident. In the lymph-nodes of three patients, the cytoplasm of mononucleate cells exhibited groups of spherieal particles, approximately 150 A in diameter, whieh eoiisisled of an electron-dense core surrounded by a membrane. These groups ineluded 20-50 elements, sometimes showing an alveolar arrangement (Figs. 2, 3 A). One cell may even contain two or three of these clusters (Fig. 2), some of these partielcs also being observed in intercellular spaces. At no time did we find any Langerhans granules in these cells. In the hepatocytes of two of these patients we observed spherical particles of variable size (between 150 A and 220 A), generally with an electron-lucent core, scattered throughout a space limited by a membrane (Fig. 3B). Pleomorphie spherical particles, 200 A in diameter, tubular forms of the same diameter and of varying length, and double-shelled particles 420 A in diamclcr
101
(Dane particles) were observed in the sera of all six patients (Fig. 3 C).
Discussion Langerhans cells in the lymph-nodes have been so far observed in histioeytosis X (Gianotti et al. 1968), in dermatopathie lymphadenopathy (Jimbow el al. 1969) and in the normal rabbit (Kondo 1969). These eells are always present in the lymph-nodes of PAC, with Langerbans granules usually always rod-shaped and never attached to the cellular membranes. The partiele aggregates found in the cytoplasm of monocytc lymphnode cells are not glycogen clusters, sinee these are totally electron-dense and lack a limiting membrane. Fhc spherical particles we observed in the cytoplasm of hepatocytes differ from those seen in the lymph-nodes in three ways: their size ranges from 150 A to 220 A; their eleetron-lueent core; and they are not elustcred but scattered throughout areas limited by a membrane. From a morphological point of view, these particles are rather similar to those demonstrated by various authors, as reported by Gerber et al. (1974), in the nucleus and in the cytoplasm of hepatocytes of acute and chronic hepatitis. However, in order to be able to prove thai they positively are the image of the HB antigen, a comparative study with immunoclcctron microscopy will be required. The particles evidenced in the blood serum with the negative-staining technique are, on the other hand, typical of the HB antigen. These findings are now conlributiiig lo the
Fig. 3 A. Particles 150 A in diameter with an electron dense core, in a lymph-node of PAC.X 170,000. B. Spherical particles ranging in size between 150 A and 220 A in hepatocytes of PAC. X 135,000. C. The three types of parlicles associated with the hepatitis B antigen, present in the serum of a patient suffering from PAC. X 280,000.
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substantiation of the view that PAC is a clinical disease due to the primary infection of the HB antigen in childhood, acquired via oral or mueous membranes (Gianotti 1974). References Bellone, A. G., Caputo, R. & Gianotti, F. (1967) Aspetti ultrastrutturali cutanei deH'acrodermatite papulosa infantile di Crosti-Gianotti. Minerva Dermatologica 42, 278-279. Caputo, R. & Lombardi, L. (1963) Oniidri ultrastrutturali dell'epidermide umana normale. Atti dclla Accademia medica lombcirda 18, 72-77. Gerber, M., Hadziyamis, S., Vissoulis, C, Scbaffner, F., Paronetto, F. & Popper, H. (1974) Electron microscopy and immunoelectionmicroscopy of cytoplasmic hepatitis B antigen in bepatocytes. American Journal of Pathology 75, 489-496. Gianotti, F., Caputo, R. & Ranzi, T. (1968) Ultrastructural study of giant cells and Langerhans ceils granules in cutaneous lesions and lymph nodes and liver biopsies from four cases of siibacute disseminated hisliocytosis of Letter-Siwe. Arcliiv fiir Klinische und Experimentelle Dermatologie 233, 238-252. Gianotti, F. (1973) Papular acrodermatitis of
childhood. An Australian antigen disease. Archives of Diseases of Childhood 48, 794— 799. Gianotti, F. (1974) Hepatitis B antigen in papular acrodermatitis in children. British Medical Jonrnal 5924, 169-170. Jean, d., Lanibeitengbi, G., Gianotti, F., Travaglini, P. & Ranzi, P. (1967) L'epatite aciita anitterica iiell'Acroderniatite papnlosa infantile. Milan: Ganassini Publ. Jimbow, K., Sato, S. & Kukita, A. (1969) Cells containing Langerhans granules in human lymph nodes of dermatopatbic lymphadenopathy. Journal of Investigative Dermatology 53, 295-299. Kondo, Y. (1969) Macrophages containing Langerhans granules in normal lymph-nodes of the rabbit. Zcitschrift fiir Zellforschnng und Mikroskopische Anatomic 98, 506—511. Millonig, G. (1962) Furlber observations on a phosphate buffer for osmium solution. Proc. 5tli Intern. Congr. of Electron Microscopy, ed. Hieese, S. S., Jr. Philadelphia, p. 8. New York-London: Academic Press. .Address: /';•()/. F. Giatiotti Clinic of Dermatology University of Milan Via Pace 9 Milan, Italy
