Complement 9 in amyloid deposits

Figures & data

Figure 1. Immunostaining of apolipoprotein E, caspase 3, complement 3 and 9 in relation to amyloid (study cohort). (A–H) Comparison between immunohistochemistry and the enrichment of investigated proteins using the example of renal ALκ amyloidosis. Serial sections: hematoxylin eosin staining (A), Congo red staining in bright light (B), immunostaining with antibodies directed against κ-light chain (C), Congo red staining in fluorescence light (D; same section as in B), immunostaining with antibodies directed against apolipoprotein E (E), complement 9 (F), caspase 3 (G) and complement 3 (H); (I) caspase 3 immunoreactive cardiomyocytes (arrows) within amyloid deposits (asterisk). (J) Complement 9 immunoreactive cardiomyocytes (arrow head) surrounded by complement 9 immunoreactive amyloid deposits. Hematoxylin counterstain.

Table 1. Presence of positivity of the respective protein within amyloid deposits differentiated by amyloid type.

	Presence of positivity within amyloid deposits
	ApoE		Cas3		C3		C9
Positive (n)	Valid	n (%)	Valid	n (%)	Valid	n (%)	Valid	n (%)
AA	10	10 (100)	10	2 (20)	1	0 (0)	10	10 (100)
AIns	4	4 (100)	4	0 (0)	4	0 (0)	4	4 (100)
ALκ	21	19 (91)	20	3 (15)	7	4 (57)	21	21 (100)
ALλ	42	40 (95)	40	4 (10)	18	6 (33)	42	42 (100)
ATTR	35	33 (94)	35	13 (37)	22	8 (36)	35	34 (97)
Aβ	3	3 (100)	3	0 (0)	1	1 (100)	3	3 (100)
Mixed	1	1 (100)	–	–	1	1 (100)	1	1 (100)
Unclassified	2	2 (100)	2	0 (0)	1	0 (0)	2	2 (100)
Total	118	112 (95)	114	22 (19)	55	20 (36)	118	117 (99)

ApoE: apolipoprotein E; Cas3: caspase 3; C3: complement 3; C9: complement 9; valid: number of samples evaluated.

Table 2. Presence of positivity of the respective protein within amyloid deposits differentiated by organ-/tissue type.

	Presence of positivity within amyloid deposits
	ApoE		Cas3		C3		C9
Positive (n)	Valid	n (%)	Valid	n (%)	Valid	n (%)	Valid	n (%)
Heart	28	26 (93)	28	12 (43)	12	4 (33)	28	27 (96)
Kidney	21	19 (91)	21	1 (5)	3	0 (0)	21	21 (100)
Skin	14	14 (100)	14	1 (7)	6	2 (33)	14	14 (100)
Synovialis of carpal tunnel	10	10 (100)	10	1 (10)	8	1 (13)	10	10 (100)
Adipose-/connective tissue	7	7 (100)	7	0 (0)	5	0 (0)	7	7 (100)
Gastrointestinal tract	7	7 (100)	7	2 (29)	4	1 (25)	7	7 (100)
Ligament of carpal tunnel	7	6 (86)	7	5 (71)	4	4 (100)	7	7 (100)
Liver	6	6 (100)	6	0 (0)	–	–	6	6 (100)
Lung	5	4 (80)	4	0 (0)	3	3 (100)	5	5 (100)
Other	13	13 (100)	10	0 (0)	10	5 (50)	13	13 (100)
Total	118	112 (95)	114	22 (19)	55	20 (36)	118	117 (99)

ApoE: apolipoprotein E; Cas3: caspase 3; C3: complement 3; C9: complement 9; other: central nervous system, conjunctiva, corner of the mouth, Lig. anulare, lymph node, N. suralis, nasopharynx, oropharynx, urinary bladder; valid: number of samples evaluated.

Figure 2. Accordance by type of amyloidosis. The proportion of immunostained area of complement 9, apolipoprotein E, caspase 3 and complement 3 in relation to the entire amyloid deposits (i.e. 100%). Presentation according to the amyloid type. Each bar corresponds to one case.

Figure 3. Accordance by histoanatomical localization. The proportion of immunostained area of complement 9, apolipoprotein E, caspase 3 and complement 3 in relation to the entire amyloid deposits (i.e. 100%). Presentation according to the organ-/tissue-type. Each bar corresponds to one case.

Table 3. Prevalence of caspase 3-/complement 9 single positive cells differentiated by organ-/tissue type.

		Caspase 3				Complement 9
		Any positive cells	Amyloid-associated	Non-amyloid-associated		Any positive cells	Amyloid-associated	Non-amyloid-associated
Positive (n)	Valid	n (%)	n (%)	n (%)	Valid	n (%)	n (%)	n (%)
Heart	28	5 (18)	5 (18)	1 (4)	28	5 (18)	5 (18)	0 (0)
Kidney	21	5 (24)	0 (0)	5 (24)	21	5 (24)	2 (10)	5 (24)
Skin	14	3 (21)	2 (14)	1 (7)	14	2 (14)	1 (7)	1 (7)
Synovialis of carpal tunnel	10	1 (10)	1 (10)	0 (0)	10	1 (10)	1 (10)	0 (0)
Adipose-/connective tissue	7	3 (43)	3 (43)	0 (0)	7	1 (14)	0 (0)	1 (14)
Gastrointestinal tract	7	4 (57)	3 (43)	2 (29)	7	2 (29)	1 (14)	1 (14)
Ligament of carpal tunnel	7	0 (0)	0 (0)	0 (0)	7	0 (0)	0 (0)	0 (0)
Liver	6	0 (0)	0 (0)	0 (0)	6	1 (17)	1 (17)	0 (0)
Lung	4	0 (0)	0 (0)	0 (0)	5	1 (20)	1 (20)	0 (0)
Other	10	4 (40)	4 (40)	1 (10)	13	5 (39)	4 (31)	1 (8)
Total	114	25 (22)	18 (16)	10 (9)	118	23 (20)	16 (14)	9 (8)

Other: central nervous system, conjunctiva, corner of the mouth, Lig. anulare, lymph node, N. suralis, nasopharynx, oropharynx, urinary bladder; valid: number of samples evaluated.

Table 4. Prevalence of caspase 3-/complement 9 single positive cells differentiated by amyloid type.

		Caspase 3				Complement 9
		Any positive cells	Amyloid-associated	Non-amyloid-associated		Any positive cells	Amyloid-associated	Non-amyloid-associated
Positive (n)	Valid	n (%)	n (%)	n (%)	Valid	n (%)	n (%)	n (%)
AA	10	4 (40)	3 (30)	2 (20)	10	2 (20)	1 (10)	1 (10)
AIns	4	3 (75)	3 (75)	0 (0)	4	1 (25)	0 (0)	1 (25)
ALκ	20	4 (20)	3 (15)	2 (10)	21	5 (24)	4 (19)	1 (5)
ALλ	40	9 (23)	4 (10)	5 (13)	42	7 (17)	5 (12)	4 (10)
ATTR	35	2 (6)	2 (6)	0 (0)	35	6 (17)	5 (14)	1 (3)
Aβ	3	3 (100)	3 (100)	1 (33)	3	1 (33)	0 (0)	1 (33)
Mixed	–	–	–	–	1	0 (0)	0 (0)	0 (0)
Unclassified	2	0 (0)	0 (0)	0 (0)	2	1 (50)	1 (50)	0 (0)
Total	114	25 (22)	18 (16)	10 (9)	118	23 (20)	16 (14)	9 (8)

Valid: number of samples evaluated.

Figure 4. Proteomics data obtained by literature search. Occurrences of apolipoprotein E (A), complement 9 (B) and complement 3 (C) in amyloid tissue sections are depicted for amyloid and organ type. Proteomic data were reviewed and extracted from amyloidosis cases analyzed by LMD LC–MS/MS (method described in [9–11]). The total number of patients tested is indicated by data point area. Positive cases are colored/grey scaled depending on the number of spectras’ median measured, whereas negative cases are depicted by black areas. The number of spectra refers to the total number of peptide spectra identified for a protein and is considered a semiquantitative measurement of protein abundance. Proteins were identified with a >95% probability. For three cases, samples were derived from more than one organ type: *heart and gastrointestinal tract (GI-tract; not otherwise specified); **lung and skin; ***heart, subcutaneous fat aspirate (SFA) and large intestine.