General Pathology

AmyloidosisFibrinoid, Hyalin

Inst. Pathol. ,1st Med. Faculty, Charles Univ. Prague http://www1.lf1.cuni.cz/~jdusk/

Jaroslava Dušková

AmyloidosisDEF.:

disorder of protein metabolism accompanied with abnormal extracellular deposition of proteinaceous material - amyloid

Amyloid = starch like

Karl Freiherr von Rokitansky (1804-1878)

Rudolf Ludwig Karl Virchow (1821-1902).

Amyloid - history

Karl Freiherr von Rokitansky (1804-1878)

Austrian pathologist, born February 19, 1804, Königgrätz, Böhmen, Austrian Empire (now Hradec Králové, East Bohemia, Czech Republic); died July 23, 1878, Wien.

Handbuch der pathologischen Anatomie IInd Band, Wien 1842

Amyloid - history 2.

Amyloidosis – morphology Macroscopy:

small amounts – invisible larger deposits – enlarged,

firm, waxy organs

Ultrastructure & Biochemistry of Amyloid90-95% non branched fibrils diam. 10-12nm

5-10% p-component - glycoprotein + fibronectin, laminin, collagen 4

Amyloidosisconformational disease (Carrell and Lomas, Lancet, 1997)

„…arises when a constituent protein undergoes a change in size or fluctuation in shape with resultant self - association and tissue deposition“

pleated β – sheet structure

Conformational diseases (Carrell and Lomas, Lancet, 1997)

Amyloidosis Prionoses - transmissible spongiform encephalopathies (incl. m. CJD)

m. Alzheimeripleated β – sheet structure

Amyloidosis

Classification: according to the source protein

(more than 20 different identified)

according to the distribution systemic (generalised) localised

Systemic Amyloidosis - I.

AL - imunocyte dyscrasia associated light chains Ig (mostly )

„primary“

Distribution: tongue, heart, GIT, liver, spleen, kidney

Associated diseases: Plasma cell myeloma, B cell lymphoma,

Systemic Amyloidosis - II.

AA - reactive systemic amyloidosis SAA = Serum Amyloid Associated

protein „secondary“Distribution: liver, kidney, spleen, GIT, lymph nodes, bowel, adipose tissue

Associated diseases: rheumatoid arthritis, chronic infections (tb, leprosy, bronchiectasiae, osteomyelitis, IBD, neoplasms MLH , RCC

Systemic Amyloidosis - III.

senile systemic SSA 25% people over the age of 80 years (!)

-normal transthyretin TTR (prealbumin)

-mostly heart & vessels invilvement

Systemic Amyloidosis - IV.

A2 - hemodialysis associated 2 microglobulin

Hereditary AA - Familial Mediterranean Fever ATTR - Famil. polyneuropatia

transthyretin (mutated form)

Systemic Amyloidosis - complications

diminished functions of some organs, esp.

KIDNEY FAILURE

IIIrd stage Amyloid nephrosis

Localised Amyloidosis - I.

Senile cardial ATTR - transthyretin -

(structurally normal)

Senile cerebral

A - -amyloid protein

Cardiac Amyloidosis – clinical manifestations

Dilated Cardiomyopathy (predominant systolic

dysfunction) Restrictive cardiomyopathy (predominant diastolic

dysfunction)

Congestive heart failure Rhytm abnormalities Coronary insufficiency Valvular dysfunction Pericardial tamponade Enhance sensitivity to digitalis glycosides Atrial thrombosis - embolisation

Localised Amyloidosis - II.Endocrine

ACal - ca medullare gl. thyreoideae AIAPP - islets of Langerhans associated AANF - isolated atrial amyloidosis

atrial natriuretic polypeptide

Nodular tumoriform amyloid deposits (tongue, lung,larynx, skin, urinary bladder, orbita)

Clinical Diagnosis of Amyloid

Scintigraphy (in vivo)using human serum amyloid

component marked with 123J

Echocardiography (atrial amyloid)

Clinical Diagnosis of Amyloid

Biochemistrysequening DNA -hered. forms

extraction of fibrils (from a biopsy

specimen)

spectrometry

sequening of the amyloid protein

Amyloidosis – morphology Macroscopy:

small amounts – invisible larger deposits – enlarged,

firm, waxy organs

Morphological Diagnosis of Amyloid

Macroscopy

–reaction Virchow I (sol. Lugolli)

Virchow II (H2SO4)

Morphological Diagnosis of AmyloidMicroscopy:

– KONGO red (+POLARISATION!) + KMnO4

– thioflavine S,T

– crystal. violet (metachromasia)

– IMMUNOHISTOCHEMISTRY (electron microscopy)

Morphological Diagnosis of Amyloid

Materials:– GIT (stomach, duodenum rectum, gingiva)

biopsy

– kidney

– sural nerve & muscle

– fat aspiration biopsy – needle with an internal

diam. 0,7-1,2mm

Röcken Ch. Sletten K.: Amyloid in Surgical Pathology Virchows Arch., 2003, 1-26

CONGO Red synthesized by young chemist at Bayer comp. 1883 as

the first of economically lucrative direct (nod needing a mordant) textile dyes

patented by AGFA 1885(Aktiengeselschaft für Anilinfarbenfabrikation)

3 weeks after the conclusion of the West Africa Conference

to Europeans in 1885, the word Congo evoked exotic images of far-off central Africa known as The Dark Continent

the Congo red stain was named „Congo“ for marketing purposes by a German textile dyestuff company in 1885

Steensma DP: „Congo“ Red. Out of Africa? Arch. Pathol.Lab.Med.,2001, 125, 250-2

Reversibility of Amyloid

The deposits are NOT irreversible. e.g.

Hrncic R. et al: Antibody mediated resolution of light chain – associated amyloid deposits. Am.J. Pathol., 2000, 157,12369-46

Progression of generalised amyloidosis can be delayed or stopped by treatment of the underlying disease.

Röcken Ch. Shakespeare Ann: Pathology, diagnosis and pathogenesis of AA amyloidosis. Virchws Arch. , 2002, 440, 11-122

Prevention & Therapy of Amyloid

Prevention & treatment of the underlying diseases

Vaccination against β am. protein in mice diminished senile plaque formation and improved memory.

Nature Medicine, 2001, 7, 18th Jan.

A β –based experimental therapies based on degrading enzymes.

Zlokovic et al.: Neurovascular Pathways and Alzheimer Amyloid β-peptide. Brain Pathol. , 2005, 15, 78-83

Fibrinoid & Hyalin

disorders of protein metabolism

Fibrinoid Change of Collagen

vessels and connective tissue damage plasmorrhagia (leakage of plasma) deposits of Ag-AB complexes staining characteristics fibrin - like

Hyaline change

Definition (historical, descriptive): intra- or extracellular change

of homogenous rose „ glassy“ appearance

in the H&E stained histological sections

Hyaline change

Extracellular:corpus albicans, scars, hyalinoses of

serous membranes

Intracellular:Crooke cells, Mallory´ hyaline,

Russell bodies

Ultrastructure Fibrinoid - collagen fibres

surrounded by plasma proteins may be reversible

Hyalin – collagen fibres increased in thickness, changed architecture ratherstable

Hyaline change

Extracellular:corpus albicans, scars, hyalinoses of

serous membranes

Intracellular:Crooke cells, Mallory´ hyaline,

Russell bodies

Significance of Fibrinoid Change diminished quality of the collagen

( firmness, permeability) tendency to thrombosis in the

vessels, aneurysms formation

Significance of Hyalin Change diminished quality of the

collagen ( elasticity) ischemia in organs with

thickened arterial walls intracellular - function, death