Differential staining of skin-limited amyloid and colloid bodies with immunofluorescence after pretreatments

We examined whether immunoglobulin (Ig) and complement (C) components of amyloid and colloid bodies were inherent parts of these substances or were present due to nonspecific absorption only. In direct immunofluorescence (IF) studies without pretreatment, skin-limited and systemic amyloid and colloid bodies in all cases showed positive staining for Ig or C. When these sections were pretreated with 0.1 M glycine buffer (pH 7.2) or 0.05% Tween-20 solution, Ig and C in skin-limited amyloid deposits were negative or weakly positive. In contrast, positive fluorescence of colloid bodies and systemic amyloid masses was not influenced by pretreatment. Existence of amyloid masses before and after pretreatment were confirmed by Thioflavin-T and Dylon stains. In addition, pretreatment did not alter disulfide bonds by DACM staining or the reactivities of amyloid with monoclonal antikeratin antibody EKH4. These results suggest that skin-limited amyloid can be differentiated from systemic amyloid or colloid bodies by these methods. We can infer from the present studies that most of the Ig and C in skin-limited amyloid masses are a result of nonspecific absorption due to penetration of serum, which is different from Ig in systemic amyloid and colloid bodies in as much as in these conditions immunobinding is specific.     

Amyloidogenesis in a case of cutaneous amyloidosis: an electron microscopic study using the refixation-reembedding method

A case of the hyperkeratotic type of macular amyloidosis was electron microscopically examined using the refixation-reembedding method. All the cytoid bodies seen in the dermis were amyloid islets associated with malformed basal lamina-like material. The uppermost amyloid islets were in close apposition to the basal cells, which had neither hemidesmosomes nor intact basal lamina. Anchoring fibril-like filaments were seen between the amyloid islets and the basal lamina-like material. Many fibrillar bodies and clusters were present in the spinous layer, but not in the dermis. Masses of eosinophilic, PAS-positive ovoid bodies seen in the horny layer were tightly packed fibrillar bodies shifted from the spinous layer to the horny layer. These findings suggest that the disturbed basal cells produce an amyloid substance in the dermis, as well as malformed basal lamina. The colloid bodies formed by the degeneration of the basal cells seemed to shift up to the horny layer but not to contribute to the amyloidogenesis in this case.     

Papillomavirus-infected keratinous cyst on the sole. A histologic, immunohistochemical, and electron microscopic study.

A 17-year-old boy had a keratinous cyst on the sole. The keratinous cyst and its overlying epidermis had solitarily scattered keratinocytes, which contained a peculiar intracytoplasmic inclusion body above the lower spinous layer. Immunohistochemistry and electron microscopy revealed that the nuclei of these cells had virions of papillomavirus. These virions appeared above the spinous layer. The inclusion bodies were highly eosinophilic masses in the viable layer, and slightly basophilic, fine granules in the cornified layer. Their number at any time was usually one. Their histologic and ultrastructural features and their N-(7-dimethylamino-4-methylcoumarinyl)-maleimide staining property were different from those of keratohyalin and from those of amyloid or hyaline body. Also the keratinous cyst was associated with colloid bodies, showing the lamellated figure in the subepithelial area.     

Colloid-amyloid bodies in PUVA-treated human psoriatic patients.

In 4 cases of PUVA-treated psoriatic patients a number of colloid bodies and amyloid-like deposits were seen at the dermo-epidermal junction, in a very similar fashion the colloid bodies and the amyloid substances are deposited in lichen planus and in primary cutaneous anyloidoses respectively. In some instances such a deposition was found within the epidermis and contained remnants of cellular debris. Serial sections revealed that a part of the intraepidermal deposit is located in the dermis. Some deposition is partially composed of typical amyloid filaments and partially of nonamyloidal filaments. The latter were identical to those found in colloid or Civatte bodies of lichen planus and other conditions. Follow-up biopsies done in 1 patient several months after the cessation of the treatment still showed these substances. It was suggested that the amyloid-like substances are the product of special degeneration (apoptosis) of epidermal cells as colloid substances: Initally these cells undergo filamentous or colloid degeneration and they drop off into the dermis; where some of the characteristics of filaments are modified and connective tissue elements are added to produce such filamentous and amorphous components as seen in the amyloid island, whereas others remain as colloid bodies. Absorption or elimination of these substances seem to be extremely slow.     

Histogenesis of primary localized cutaneous amyloidosis: sequential change of epidermal keratinocytes to amyloid via filamentous degeneration.

Two cases of lichen amyloidosus and 8 cases of macular amyloidosis were examined by electron microscopy. Epidermal keratinocytes showed variable degrees of focal degeneration in the basal or lower Malpighian layer. The primary change was seen in cells which contain fibrillar (30 nm in thickness) cytoplasmic inclusion. The following developments seemed to lead to filamentous degeneration (colloid bodies): (1) aggregation of tonofilaments within the granular or fibrillar cytoplasm, (2) filamentous cells, which are composed of bundles of 7-nm thick filaments surrounded by cell membrane and desmosomes, and (3) filamentous masses composed of bundles or whorls of tightly packed 7-nm thick filaments in the intercellular spaces. At the dermo-epidermal junction, some of the filamentous masses were surrounded by the basal lamina of the epidermis and others were dropping into the dermis. Occasionally, loosened filaments (similar to amyloid filaments) were about to drop into the dermis. Early formation of amyloid islands consisted of electron-dense and electron-light parts. They were located directly beneath the epidermis. In the deeper postion of the papillary dermis and in the upper reticular dermis, the majority were electron-light masses. Electron dense parts were the densely packed 7-nm thick filaments, whereas electron-light parts were the typical straight amyloid filaments. Small tubular filaments were seen in common in the filamentous cells, filamentous masses, and amyloid islands. It is concluded that some of the amyloid substance in primary localized forms of cutaneous amyloidoses derive from the epidermal cells through filamentous degeneration.     

A case of lichen amyloidosus with Riehl's melanosis-like lesion on the face: a histological and electron microscopic study of Civatte body and amyloid.

Ultrastructural studies were carried out on the patient who developed lichen amyloidosus and Riehl's melanosis-like lesion on the legs and face, respectively. In the skin specimens from the face, there were many masses of aggregations of wavy or net-like filaments which appeared to correspond to Civatte bodies as recognized under the light microscope at the dermo-epidermal junction. Sometimes, an other kind of filament mass was present in the papilla. These were aggregations of straight and non-branching filaments and were very similar to so-called amyloid masses. These amyloid-like masses, which seemed to be in a developmental stage, consisted of net-like filaments as well as straight and non-branching filaments. Therefore, the net-like filament masses or Civatte bodies seemed to serve a particular role in the formation of these amyloid-like filament masses. On the other hand, in the skin specimens from the leg, there were many amyloid masses in the papillae and few Civatte bodies in the dermo-epidermal junction, but no features suggesting that Civatte bodies may serve a role in the formation of amyloid. From these observations, it may be concluded that Civatte bodies will contribute to the formation of amyloid under certain conditions like Riehl's melanosis; however, it is still uncertain whether Civatte bodies also contribute to the formation of amyloid filaments of lichen amyloidosus.     

Lichen planus: an ultrastructural study.

Ultrastructural features of skin lesions are described in 11 patients with lichen planus. Keratinocytes and melanocytes in the basal layer show loss of nuclear membrane, disappearance of nucleoli, homogenization of nuclear material, and aggregation of tonofilaments around the nucleus to resemble cells in the late prophase of the mitotic cycle except that there is no clumping of chromosomes. These cells undergo fibrillar transformation to form densely packed fibrillar bodies (colloid bodies) which are the size and shape of normal cells and frequently have cytoplasmic organelles. Ten per cent of the mainly lympho-histiocytic cells in the dermal infiltrate show changes such as multiple cell membrane discontinuities, disintegration of cytoplasm, and breaks in the nuclear membrane with spilling of nuclear substance either into the cytoplasm or the extracellular space. The shape, size and occasional fibrillar changes in the nuclei and cytoplasm of these cells resemble colloid bodies. Contact sites frequently occurred between cells in the dermal infiltrate, especially between lymphocytes and macrophages. It appears that primary injury to the basal layer cells occurs during the early phase of the mitotic cycle and antigen from this primary site may evoke a cell-mediated type of hypersensitivity reaction. Colloid bodies evolve from cells which have injured nuclei and undergo fibrillar transformation. Dermal as well as epidermal cells may contribute to colloid body formation.     

Nodular colloid degeneration: distinctive histochemical and ultrastructural features

We recently evaluated a patient with a unique form of nodular colloid degeneration. Lesions consisted of pruritic, purpuric plaques involving the upper and lower extremities and the neck. Examination of a biopsy specimen revealed large, fissured masses of amorphous material in addition to solar elastosis. The amorphous material showed negative reactions to stains for amyloid, and other histochemical studies showed results similar to those for colloid milium. The amorphous masses demonstrated a marked positive reaction to the Verhoeff-van Gieson stain for elastin. Electron microscopic examination revealed masses of degenerated elastic tissue bordered by fibrillar material that resembled amyloid morphologically. We believe this case represents a nodular form of solar degeneration with clinical and histologic resemblances to both colloid milium and cutaneous amyloidosis.     

Colloid body formation in bullous pemphigoid

Summary Thirty-eight biopsy specimens from 18 cases of bullous pemphigoid (BP) were observed using direct immunofluorescence (IF) techniques with fluorescein isothiocyanate (FITC)-labelled antisera against human serum factors. In addition to deposits of immunoglobulins and serum components at the basement-membrane zone (BMZ), 15 specimens from eight cases displayed homogeneous and globular fluorescent bodies in the uppermost dermis and/or the blisters when FITC-labelled antisera to human IgM and other serum factors were used. Using immunoperoxidase staining, haematoxylin/eosin (HE) and periodic acid-Schiff (PAS) staining, these immunoglobulin and/or complement-positive cell-sized bodies were shown to be slightly eosinophilic and PAS positive. Electron microscopy revealed entangled networks of microfilaments approximately 7–8 nm in diameter. These homogeneous, fibrillar bodies were histologically, immunohistologically and ultrastructurally indistinguishable from the colloid bodies found in lesional skins of lichen planus, lupus erythematosus, dermatomyositis and several other dermatoses. In BP, degenerated keratinocytes adjacent to the blister roof, may, after undergoing a filamentous change, drop off into the dermis and subsequently form homogeneous, fibrillar bodies in the uppermost dermis when reepithelization is completed.     

Non‐Hodgkin's lymphoma and auricular hypoplasia: associated with juvenile colloid milium or ligneous conjunctivitis?

The exact nature of amyloid‐like hyaline material deposits in the skin is not well understood in some disorders. Three of those – ligneous conjunctivitis, ligneous periodontitis and colloid milium – have been rarely reported in a same patient. We report a case of mucosal and skin deposits of an amyloid‐like homogeneous material associated with non‐Hodgkin's lymphoma and congenital auricular hypoplasia. We discussed and reviewed the literature on these unique associations to determine whether these are the same pathological process. We also noted whether this case represents a new syndrome or a coincidental association.     

Nodular cutaneous amyloidosis. Clinical, histopathological and ultrastructural findings]

A case of nodular amyloidosis cutis of the face with minimal deposits of amyloid in the rectum is reported with respect to clinical, histopathological, and ultrastructural findings. A discussion of the problem of systematization in nodular amyloidoses is included. By electron microscopy, the principal similarity of the ultrastructure of amyloid is stressed again. Amyloid in this case of nodular amyloidosis cutis is thought to be synthesized by plasma cells which are found especially in the growing peripheral parts of the tumors. Fibroblasts with extremely dilated endoplasmic reticulum are almost totally surrounded by the fibrillar masses of amyloid within the tumor. Unusual spindle-formed cisternae of Golgi bodies containing oriented coarse fibrils are found in the fibroblasts. They are thought to represent a disturbance of the secretion of tropocollagen to the interstitium due to the surrounding amyloid masses.     

Sulphhydryl crosslinking in cutaneous apoptosis: A review

Histochemical findings of cutaneous apoptosis are reviewed with special reference to the sulphhydryl groups. Using stain techniques with the new fluorescent thiol reagent, N-(7–dimethylamino-4–methylcoumarinyl) maleimide (DACM), it was shown that sulphhydryl crosslinking occurs in colloid bodies and sunburn cells and also participates, at least in part, in amyloid formation. As the DACM staining clearly demonstrated the localization of sulphhydryl and disulphide groups in the epidermis, this technique can be of help dealing with the problem of the origin of cutaneous apoptotic bodies.     

Rippled-pattern sebaceoma.

A 71-year-old woman had a dome-shaped, slightly erythematous nodule on the anterior scalp. The nodule histopathologically revealed sebaceoma based on the silhouette and cytology. A notable and unique finding was often observed in the aggregations of sebaceoma; an arrangement of small, monomorphous, cigar-shaped basaloid cells in linear rows parallel to one another, resembling the palisading of nuclei of Verocay bodies, namely a rippled-pattern. Although we are not certain that sebaceoma can be clearly separated from trichoblastoma with sebaceous differentiation in all cases, in the present case, the absence of an abundant and densely fibrotic stroma, of follicular differentiation, and of a palisading border in the neoplastic aggregations as well as the presence of many vacuolated cells and tiny duct-like spaces favors the diagnosis of sebaceoma rather than trichoblastoma with sebaceous differentiation. Based on the expression patterns of CKs as well as similar cytological features between germinative cells in our case and immature cells in the mantles of normal vellus follicles, we believe that rippled-pattern sebaceoma is composed of immature sebaceous germinative cells with some foci of advanced sebaceous differentiation (toward the sebaceous duct and sebaceous lobule).     

Colloid milium: light and electron microscopic investigations

A case of colloid milium (‘juvenile type’) is presented. Histochemically the colloid material showed the well known similarity to amyloid where especially in the upper portions of the colloid immunoglobulin and complement depositions could be found. Fibrin was demonstrated predominantly in the periphery of individual colloid islands. Electron microscopically, colloid exhibited a fibrillary structure. On the surface of the islands as well as enclosed within the colloid material numerous desmosome-like structures could be visualized and sometimes also nuclear remnants were present. In circumscribed areas a fibrillary transformation of keratinocytes was observed. On the basis of these findings it is concluded that colloid represents tonofilament-like material and therefore is of epidermal origin.     

Juvenile colloid milium

A 7-year-old Italian girl with juvenile colloid milium was studied with histological, immunohistochemical, and electron microscopic methods. This patient had a well-documented history of severe sunburn and developed the lesions on the face shortly afterward. Numerous apoptotic keratinocytes were observed in the lower epidermis. These cells began their degeneration with filamentous whorl formation (or filamentous degeneration) of tonofilaments. In the papillary dermis the colloid substance was resolved by the electron microscopy into either wavy, thin filaments derived from the epidermal keratinocytes or typical amyloid filaments. Many desmosomes and gap junctions were found in the colloid substance. Polyclonal antikeratin antibody (DAKO) was positive in the colloid substance, particularly in the parts close to the epidermis. These findings suggested that juvenile colloid milium is different from adult colloid milium despite clinical similarities and that the former belongs to the group of actinic amyloid K, i.e. amyloidoses due to actinic degeneration of keratinocyte and its keratin.     

Vitiligo with an inflammatory erythema in Vogt-Koyanagi-Harada disease: demonstration of filamentous masses and amyloid deposits

A 35-year-old Japanese woman developed vitiligo with an inflammatory erythema on the face two years after the onset of bilateral uveitis and dysacusis. Histological studies of the lesions revealed the loss of pigment and dense mononuclear cell infiltration around the blood vessels. At the dermoepidermal junction area, a few eosinophilic masses were found. Electron microscopy disclosed that the masses were filamentous masses (colloid bodies) and amyloid. After the oral administration of prednisolone, the erythema subsided, and the vitiligo showed some regimentation.     

Sulphhydryl and disulphide stainings of subepidermal hyaline bodies

The localization of sulphhydryl (-SH) groups and disulphide (-S-S-) bonds in subepidermal hyaline bodies was examined using histochemical techniques with the specific fluorescent thiol reagent (DACM). In lichen planus, hyaline bodies showed -S-S- but not -SH staining, suggesting that erythematosus and dermatomyositis, there were not only -S-S-positive hyaline bodies, considered of epidermal origin, but also -S-S- negative ones. The latter may consist of dermal components, since no dermal components are stainable with DACM. DACM staining therefore seems to be of help in determining the origin of hyaline bodies.     

Lichenoid tissue reaction

Lichenoid tissue reaction (LTR) is characterised by epidermal basal cell damage which takes the form of liquefaction degeneration or cell death either apoptosis or necrosis with an associated cascade of histologic events in epidermis and dermis. LTR is found in clinical conditions with lichenoid poikilodermatous and pigmentary dermatoses. A selected group of fifty lichenoid and pigmentary dermatoses such as Lichen planus (LP) Discoid lupus erythematosus (DLE) Lichenoid melanodermatitis (LM) and Lichen nitidus (LN) were studied. In LP basal cell liquefaction degeneration was extensive in comparison to other disease with large number of Civatte bodies and colloid bodies. There were significant vasodilatation in upper dermis inside the massive band like infiltrate. PAS positive basement membrane was disrupted in reaction area. Hypergranulosis was conspicuous. Chronic DLE showed spotty lichenoid reaction in the form of basal cell liquefaction degeneration. Civatte bodies and colloid bodies were infrequent. Infiltrate was more focal but could be band like. Epidermal atrophy and thickening of PAS positive basement membrane were important differentiating features, LM or Melanodermatitis toxica revealed focal mild to moderate liquefaction degeneration of basal cells with atrophy of the epidermis. The infiltrate although band like was less dense with marked pigmentary incontinence in clumps and giant melanophages. Civatte bodies, colloid bodies were not found and vascular changes were less prominent. LN showed localised basal cell damage with claw like rete ridges clutching a dense infiltrate. The dermal infiltrate often showed multinucleated giant cell. Civatte bodies and colloid bodies were not present. In some cases of the overlap syndrome LP/LE a careful study of lichenoid tissue reaction could distinguish these two diseases.(ABSTRACT TRUNCATED AT 250 WORDS)     

Half-and-half cells in lichen planus

In the course of a study of wound healing in four patients with lichen planus, we found transformed keratinocytes with a hitherto undescribed ultrastructure in both wounded and undisturbed papules. We have called these epidermal cells half-and-half cells because they showed changes on the one hand of increased synthetic activity and, on the other hand, of fibrillar transformation closely resembling that seen in fully developed colloid bodies. We suggest that these half-and-half cells may provide a useful clue to the early changes occurring in colloid body formation.