図 1-7.  各蛋白質における小松川株と RV303 株のアミノ酸配列の比較

各蛋白質上の数字は，小松川株と RV303 株の間で異なったアミノ酸の位置を示す。その位置における小 松川株および RV303 株のアミノ酸を一文字表記により示した。

   

COOH COOH

COOH

COOH

COOH H2N

60 65 82 160 172 284

H2N

80

N 蛋白質

L F

D E G S R Q

N V

R K

N S

H2N

155

T I

156

R K

H2N

-5

S P

187

T A

478

G E

H2N

118 T

334

N A V

597

V I

1,124

K L

2,085

A T 100 aa

200 aa 小松川

RV303

小松川 RV303

小松川 RV303

小松川 RV303

小松川 RV303

450 aa

297 aa P 蛋白質

202 aa M 蛋白質

G 蛋白質

L 蛋白質 505 aa

2,127 aa

: N, P, M, G 蛋白質

: L 蛋白質

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

MDADKIVFKVNNQVVSLKPEIIVDQYEYKYPAIKDLKKPSITLGKAPDLNKAYKSVLSGLNAAKLDPDDVCSYLAAAMQF ......L ...... ...C....M...... ...C....M...... ....R...RS...R....A...C....M...... ..S....C....M...... ...C....MS..... FEGTCPEDWTSYGILIARKGDKITPDSLVEIKRTDVEGNWALTGGMELTRDPTVPEHASLVGLLLSLYRLSKISGQNTGN ...... ...... ...S....... ...S....... ...N....N....M....S...... ...R...N....S....... ...V....G...... YKTNIADRIEQIFETAPFIKIVEHHTLMTTHKMCANWSTIPNFRFLAGTYDMFFSRIEHLYSAIRVGTVVTAYEDCSGLV ...... ...... ...V.V...... ...V...... ...V...... ...V...... ...V...... SFTGFIKQINLTAREAILYFFHKNFEEEIRRMFEPGQETAVPHSYFIHFRSLGLSGKSPYSSNAVGHVFNLIHFVGCYMG ...... ...... ...G...... ...... ...... ...... ...... QVRSLNATVIAACAPHEMSVLGGYLGEEFFGKGTFERRFFRDEKELQEYETAELTKTDVALADDGTVNSDDEDYFSGETR ...... ...... ...T...A....... ...A....... .I...R...A........D....A. ...A.....S....... ...A....... SPEAVYARIMMNGGRLKRSHIRRYVSVSSNHQARPNSFAEFLNKTYSSDS? ...? ...I...? ...T....? ...T....N..? G...? ...T....N..? ...T....?

1020304050607080 90100110120130140150160 170180190200210220230240 250260270280290300310320 330340350360370380390400 410420430440450

図 1 -8 .  N 蛋 白 質 の ア ミ ノ 酸 配 列 の 比 較

小松川株と同じアミノ酸をドットで表記した。 赤枠: 小松川株とRV303株で異なるアミノ酸残基、青枠: 病原性に重要な既知のアミノ酸残基 AR: 系統群Arctic-Related 、Cosmo: 系統群Cosmopolitan

RNA結合領域273 394395

  

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

1020304050607080 90100110120130140150160 170180190200210220230240 250260270280290

図 1 -9 .  P 蛋 白 質 の ア ミ ノ 酸 配 列 の 比 較

L蛋白質結合領域 N蛋白質結合領域

可変領域 可変領域

系統群Arctic-Related 街上毒 系統群Cosmopolitan 街上毒 固定毒系統群Cosmopolitan 小松川株と同じアミノ酸をドットで表記した。 赤枠: 小松川株とRV303株で異なるアミノ酸残基、青枠: 病原性に重要な既知のアミノ酸残基

65 82 172 284

66 81 226

5658

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

MNFLRKIVKNCRDEDTQKPSPVSAPPDDDDLWLPPPEYVPLKELTGKKNMRNFCINGEVKVCSPNGYSFRILRHILRSFD ....................................................................... ...I................................................................... ...............P.........................SR.........E.................. ...............A.........................S...I......................... ..I........K.........A.................F.S............................. ..V......K................Y..............S........V.....A...............N .....................A...L....P........P.S...........G...............K... EIYSGNHRMIGLVKVVIGLALSGAPVPEGMNWVYKLRRTLIFQWADSRGPLEGEELEYSQEITWDDDTEFVGLQTRVSAR ...................................................................IK.... ...................................................................I... ...............................................................A......I... ...................................................................I... .....................................................H.............I... ................V.................................................I... .......................S............F...............................I..I.K QCHIQGRIWCINMNSRACQLWSDMSLQTQRSEEDKDSSLLLE* ....................................... ....................................... ....................................... ....................................... ............T.............K..M......... ............T.......................... ..............P........................

1020304050607080 90100110120130140150160 170180190200

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

図 1 -1 0 .  M 蛋 白 質 の ア ミ ノ 酸 配 列 の 比 較

_N^{蛋白質結合領域}

街上毒 街上毒 固定毒

Arctic-Related Cosmopolitan Cosmopolitan 小松川株と同じアミノ酸をドットで表記した。 赤枠: 小松川株とRV303株で異なるアミノ酸残基、青枠: 病原性に重要な既知のアミノ酸残基

95155156

MVPQALLFVPLLVFPMCFGKFPIYTIPDKLGPWSPIDIHHLSCPNNLVVEDEGCTNLSGFSYMELKVGYISAIKVNGFTC ..............S................................................................. ..............S................................................................. .......V....G.SL................................................................ ....V.......A.SL................................................................ .......L..I.G.SS............T................................................... ....V....L..G.SL............E.............................E..................... ...............L......................................................L...M..... TGVVTEAETYTNFVGYVTTTFKRKHFRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVKTTKESLVIISPSVAD ................................................................................ ................................................................................ ...............................E................................................ ................................................................................ ....................................................S........................... ................................................................R......I......T. ...................................................Q......R..................... LDPYDKSLHSRVFPNGKCSGITISSTYCPTNHDYTIWLPENPRLGASCDIFTNSRGKRASKGSKTCGFVDERGLYKSLKG .............................................T.................................. .............................................T.................................. ..............S.......V.....S........M.......T.................................. ..............G.......V.....S........M.......T.................................. .....N........S.......V..V..S......V.M..SL...T.................................. ..............G.......V.....S........M.....PRTP.............N.N................. .....R........S.....VAV.....S........M.......M.................E................ ACKLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHDFRSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRL ................................................................................ ................................................................................ ............K........................................D.......................... ................................................................................ .........................N...............L.........I.................I.......... ..R.......................K..................................................... .........................N................H.............R....................... SHLRKLVPGFGKAYTIFNKTLMEADAHYKSVRTWNEIIPSKGCLRVGGRCHPHVNGVFFNGIILGPDGHVLIPEMQSSLL ................................................................................ ................................................................................ ....................................V.......K................V.................. ................................................................................ .Y......................E....................................................... ............................................K......................D............ ....................................................................N........... QQHMELLESSVIPLMHPLADPSTVFKDGDEAEDFVEVHLPDVHKQISGVDLGLPNWGKYVLVSAGVLIALMLIIFLMTCC ................................................................................ ................................................................................ R............................................V..............SL...A......M....... .............................................V...............LG..A......M....... ...I.................F........T...I........E.V...........E...L...T.............. ..........................E...............Y..................MT..AM.G.V..FS...W. ..............V........I...................N.V...............L...A.T............ GRVNRPKSTQRNLGGTERKVSVTSQSGKVISSWESYKSGGETRL? ................G...........................? ..........H.....G...........................? RK....EP...S.E.KG.........................S.? R.....E....S....G....I......................? RK.D..E....S.R..G.N.........F.P...........G.? R.A...E.K..SF...GGN...........P..........I..? R....SEP..H..R..G.E....P....I......H........?

株 株  株 株 -Dog 株 株 S 株 A 株 株 株  株 株 -Dog 株 株 S 株 A 株 株 株  株 株 -Dog 株 株 S 株 A 株 株 株  株 株 -Dog 株 株 S 株 A 株 株 株  株 株 -Dog 株 株 S 株 A 株 株 株  株 株 -Dog 株 株 S 株 A 株 株 株  株 株 -Dog 株 株 S 株 A 株

102030405060 708090100110120130140 150160170180200210220 230240250260270280290300 310320330340350360370380 390400410420430440450460 470480490500

1シグナルペプチド細胞外領域

図 1 -1 1 .  G 蛋 白 質 の ア ミ ノ 酸 配 列 の 比 較

膜貫通領域 細胞内領域 小松川株と同じアミノ酸をドットで表記した。 赤枠: 小松川株とRV303株で異なるアミノ酸残基、青枠: 病原性に重要な既知のアミノ酸残基 緑枠: 糖鎖付加に必要なアミノ酸配列、オレンジ枠: 推定のニコチン性アセチルコリン受容体結合部位

-5 187 478

242255 333

37 204 247 319

147189214 268

190

  

KKVDLGSLKVGGTAAQSMISLWLYGAHSESNRSRKCITELAHFYSKSSPIEKLLNCTLGNRGLRIPPEGVLSCLERVDYD .....................................N.......................................... ......................S..............I....................................D..... ..................................R...D......................................... ..................................R...D......................................... ..................................R...D....................................K.... ..................V....C..........R...D................................N.....N.. ............M.....................R...D................L........................ HSLGDFPAFIKDKASQLEGTFGPSAKRFFGVLDQFDNIHDLVFVYGCYRHWGHPYIDYRKGLSKLYDQVHIKKVIDKSYQ .............V.................................................................. ....G........V.................................................................. .......V.....V............K..K.................................................. .......V.....V...............R.................................................. .......V.....V...............K........................................M......... .P.....M.....VN..............R.................................................. .......V.....V....E....C.R...RA................................................. LRLYFVITEKLLANYILPLFDALTMTDNLNKVFKKLVDRVTGQGLLDYSRVTYAFHLDYEKWNNHQRLESTEDVFSVLDQ ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... RTKILAQGDNQVLCPTYMLSPGLSREGLLYELESISRNALSIYRAIEEGASKLGLIIKKEETMCSYDFLIYGKTPLFRGN ................................................................................ ................................................................................ ................................................................................ ................................................................................ ........................L....................................................... ........................Q....................................................... ........................Q........R...........V.................................. MLGFLPKSSISCSCGATGGGNPRVSVSVLPSFDQSFFSRGPLKGYLGSSTSMSTQLFHAWEKVTNVHVVKRALSLKESIN ...L............................................................................ ...L............................................................................ ...L........T...I............................................................... ...L........T................................................................... ...L........T................................................................... ..ELF.......T................................................................... ...L........T................................................................... KKIIVVKKAATTRRLLVVGGSSIIEEDDLLSSGGEEVVEERRHHLLRRGGYYNNRRWWIITLEDIRSRSSLLDYSCL ....T.......................................... ....T.............I............................ ....T......A...........H....................... ....T..................H....................... ....T.....N.....R......H........K.............. ....T...I...K..........H....................... ....T.......K...R......HR......................

KKVDLGSLKVGGTAAQSMISLWLYGAHSESNRSRKCITELAHFYSKSSPIEKLLNCTLGNRGLRIPPEGVLSCLERVDYD .....................................N.......................................... ......................S..............I....................................D..... ..................................R...D......................................... ..................................R...D......................................... ..................................R...D....................................K.... ..................V....C..........R...D................................N.....N.. ............M.....................R...D................L........................ HSLGDFPAFIKDKASQLEGTFGPSAKRFFGVLDQFDNIHDLVFVYGCYRHWGHPYIDYRKGLSKLYDQVHIKKVIDKSYQ .............V.................................................................. ....G........V.................................................................. .......V.....V............K..K.................................................. .......V.....V...............R.................................................. .......V.....V...............K........................................M......... .P.....M.....VN..............R.................................................. .......V.....V....E....C.R...RA................................................. LRLYFVITEKLLANYILPLFDALTMTDNLNKVFKKLVDRVTGQGLLDYSRVTYAFHLDYEKWNNHQRLESTEDVFSVLDQ ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... RTKILAQGDNQVLCPTYMLSPGLSREGLLYELESISRNALSIYRAIEEGASKLGLIIKKEETMCSYDFLIYGKTPLFRGN ................................................................................ ................................................................................ ................................................................................ ................................................................................ ........................L....................................................... ........................Q....................................................... ........................Q........R...........V.................................. MLGFLPKSSISCSCGATGGGNPRVSVSVLPSFDQSFFSRGPLKGYLGSSTSMSTQLFHAWEKVTNVHVVKRALSLKESIN ...L............................................................................ ...L............................................................................ ...L........T...I............................................................... ...L........T................................................................... ...L........T................................................................... ..ELF.......T................................................................... ...L........T................................................................... KIVKATRLVGSIEDLSGEVERHLRGYNRWITLEDIRSRSSLLDYSCL ....T.......................................... ....T.............I............................ ....T......A...........H....................... ....T..................H....................... ....T.....N.....R......H........K.............. ....T...I...K..........H....................... ....T.......K...R......HR......................

VGGTAAQSMISLWLYGAHSESNRSRKCITELAHFYSKSSPIEKLLNCTLGNRGLRIPPEGVLSCLERVDYD ............................N.......................................... .............S..............I....................................D..... .........................R...D......................................... .........................R...D......................................... .........................R...D....................................K.... .........V....C..........R...D................................N.....N.. ...M.....................R...D................L........................ IKDKASQLEGTFGPSAKRFFGVLDQFDNIHDLVFVYGCYRHWGHPYIDYRKGLSKLYDQVHIKKVIDKSYQ ....V.................................................................. ....V.................................................................. ....V............K..K.................................................. ....V...............R.................................................. ....V...............K........................................M......... ....VN..............R.................................................. ....V....E....C.R...RA................................................. KLLANYILPLFDALTMTDNLNKVFKKLVDRVTGQGLLDYSRVTYAFHLDYEKWNNHQRLESTEDVFSVLDQ ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... NQVLCPTYMLSPGLSREGLLYELESISRNALSIYRAIEEGASKLGLIIKKEETMCSYDFLIYGKTPLFRGN ....................................................................... ....................................................................... ....................................................................... ....................................................................... ...............L....................................................... ...............Q....................................................... ...............Q........R...........V.................................. ISCSCGATGGGNPRVSVSVLPSFDQSFFSRGPLKGYLGSSTSMSTQLFHAWEKVTNVHVVKRALSLKESIN ....................................................................... ....................................................................... ...T...I............................................................... ...T................................................................... ...T................................................................... ...T................................................................... ...T................................................................... GSIEDLSGEVERHLRGYNRWITLEDIRSRSSLLDYSCL ...................................... .........I............................ ..A...........H....................... ..............H....................... .N.....R......H........K.............. ...K..........H....................... ...K...R......HR......................

KKVDLGSLKVGGTAAQSMISLWLYGAHSESNRSRKCITELAHFYSKSSPIEKLLNCTLGNRGLRIPPEGVLSCLERVDYD .....................................N.......................................... ......................S..............I....................................D..... ..................................R...D......................................... ..................................R...D......................................... ..................................R...D....................................K.... ..................V....C..........R...D................................N.....N.. ............M.....................R...D................L........................ HSLGDFPAFIKDKASQLEGTFGPSAKRFFGVLDQFDNIHDLVFVYGCYRHWGHPYIDYRKGLSKLYDQVHIKKVIDKSYQ .............V.................................................................. ....G........V.................................................................. .......V.....V............K..K.................................................. .......V.....V...............R.................................................. .......V.....V...............K........................................M......... .P.....M.....VN..............R.................................................. .......V.....V....E....C.R...RA................................................. LRLYFVITEKLLANYILPLFDALTMTDNLNKVFKKLVDRVTGQGLLDYSRVTYAFHLDYEKWNNHQRLESTEDVFSVLDQ ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... ....................................I........................................... RTKILAQGDNQVLCPTYMLSPGLSREGLLYELESISRNALSIYRAIEEGASKLGLIIKKEETMCSYDFLIYGKTPLFRGN ................................................................................ ................................................................................ ................................................................................ ................................................................................ ........................L....................................................... ........................Q....................................................... ........................Q........R...........V.................................. MLGFLPKSSISCSCGATGGGNPRVSVSVLPSFDQSFFSRGPLKGYLGSSTSMSTQLFHAWEKVTNVHVVKRALSLKESIN ...L............................................................................ ...L............................................................................ ...L........T...I............................................................... ...L........T................................................................... ...L........T................................................................... ..ELF.......T................................................................... ...L........T................................................................... KIVKATRLVGSIEDLSGEVERHLRGYNRWITLEDIRSRSSLLDYSCL ....T.......................................... ....T.............I............................ ....T......A...........H....................... ....T..................H....................... ....T.....N.....R......H........K.............. ....T...I...K..........H....................... ....T.......K...R......HR......................

VGGTAAQSMISLWLYGAHSESNRSRKCITELAHFYSKSSPIEKLLNCTLGNRGLRIPPEGVLSCLERVDYD ............................N.......................................... .............S..............I....................................D..... .........................R...D......................................... .........................R...D......................................... .........................R...D....................................K.... .........V....C..........R...D................................N.....N.. ...M...................................RR......DD...........................L........................ IKDKASQLEGTFGPSAKRFFGVLDQFDNIHDLVFVYGCYRHWGHPYIDYRKGLSKLYDQVHIKKVIDKSYQ ....V.................................................................. ....V.................................................................. ....V............K..K.................................................. ....V...............R.................................................. ....V...............K........................................M......... ....VN..............R.................................................. ....V....E....C.R...RA................................................. KLLANYILPLFDALTMTDNLNKVFKKLVDRVTGQGLLDYSRVTYAFHLDYEKWNNHQRLESTEDVFSVLDQ ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... ...........................I........................................... NQVLCPTYMLSPGLSREGLLYELESISRNALSIYRAIEEGASKLGLIIKKEETMCSYDFLIYGKTPLFRGN ....................................................................... ....................................................................... ....................................................................... ....................................................................... ...............L....................................................... ...............Q....................................................... ...............Q........R...........V.................................. ISCSCGATGGGNPRVSVSVLPSFDQSFFSRGPLKGYLGSSTSMSTQLFHAWEKVTNVHVVKRALSLKESIN ....................................................................... ....................................................................... ...T...I............................................................... ...T................................................................... ...T................................................................... ...T................................................................... ...T................................................................... GSIEDLSGEVERHLRGYNRWITLEDIRSRSSLLDYSCL ...................................... .........I............................ ..A...........H....................... ..............H....................... .N.....R......H........K.............. ...K..........H....................... ...K...R......HR......................

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株

図 1 -1 2 .  L 蛋 白 質 の ア ミ ノ 酸 配 列 の 比 較

L蛋白質において小松川株およびRV303株の間で異なるアミノ酸[図(A)①~⑤]を含む領域について、8株の狂犬病ウイルス株のアミノ酸配列を比較した[図(B)①~⑤]。 小松川株と同じアミノ酸をドットで表記した。赤枠は小松川株とRV303株で異なるアミノ酸を示す。

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株 小松川株 RV303 株 BV9901PJ 株 1088 株 RABV-Dog 株 西ヶ原株 CVS 株 ERA 株

110120130330340350 5906006101,1301,1401,150 2,0902,1002,110

ERA 株

(A ) （B ） ①

C OOH H

2

N 118 334 597 1, 124 2, 085 L  蛋白質 ①② ③ ④ ⑤ ② ③ ④ ⑤

118334 5971,124 2,085

              

   

図 1-13.  街上毒株および固定毒株の G 蛋白質上の糖鎖付加部位 

Yamada ら(74)および Hamamoto ら(17)による，街上毒株および固定毒株の G 蛋白質上の糖鎖付加部 位の数ならびに位置に関する報告に基づいて作成した。逆三角形はN型糖鎖付加部位を示す。

 

   

糖鎖付加部位の数

RV303 (KY860613) 1088 (AB645847)

RABV-Dog [5989/lib205/12] 

(LN879480) 等

2

237

SHBRV-18 (AY705373) 2

247 A11̲30558 (JX871881)

GA31940 (JQ595325) 等

2 MO1197 (KF4845541)

9704ARG (EU293116) 等

1

37 204 319

H2N COOH

CVS (LC325820) PM1503 (DQ099525)

等

3

37 319

H2N COOH

247 Nishigahara (AB044824)

ERA (GQ406342) 等

3

37 319

H2N COOH

158 LEP-Flury (GU565703)

HEP-Flury (AB085828) 3

H2N COOH

PV (M13215) 4

37158 247319 街上毒株

固定毒株 小松川

37 319

H2N COOH 2

株名 G 蛋白質

37 319

H2N COOH

319

H2N COOH

319

H2N COOH

319

H2N COOH

˜

˜ 2 —

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図 2-1. NA 細胞における小松川株，街上毒各株および固定毒各株の増殖能

各ウイルス株を MOI= 0.001 の条件で NA 細胞に接種した後，０，1，３および５日目の培養液を回収 し，各培養液中のウイルスの感染価を測定した。エラーバーは標準誤差を示す（n=3）。 

* :  有意差あり（p < 0.05），ns:  有意差なし（p  0.05  ）vs.  小松川株

   

2 3 4 5 6 7 8 9

ウイルス感染価 (Log10FFU/ml)

感染後日数

固定毒株

街上毒株

* *

*

**

*

ns

ns

*

_*

0 1 3 5

Koma ERA CVS Nishigahara

RABV-Dog 1088 西ヶ原 小松川

    

   

図 2-2. NA 細胞における小松川株，街上毒各株および        固定毒各株のフォーカス形成   

各ウイルス株を MOI= 0.0002 の条件で NA 細胞に接種後，培養 3 日目に細胞固定を実施した上で，

抗 N 蛋白質抗体を用いた蛍光免疫染色を実施した。

各ウイルス株のフォーカス像。スケールバーは 200 µm を示す。

各ウイルス株のフォーカスの面積(µm²)。ランダムに選ばれた 50 個のフォーカスの平均値 を示す。

      エラーバー:  標準誤差，* :有意差あり（p < 0.05）， 

      ns:    有意差なし（p  0.05）vs.  小松川株 

西ヶ原 CVS ERA

小松川 1088 RABV-Dog

街上毒株

固定毒株

(A)

Komatsugawa 1088 RABV-Dog Nishigahara CVS ERA

0 20000 40000 60000 80000

面積(µm2)

街上毒株 固定毒株

（B）

*

*

ns ns

ns

Koma小松川 西ヶ原1088 RABV-Dog CVS ERA

                      

表 2-1.  小松川株，街上毒各株および固定毒各株を脳内接種されたマウスの          致死率（%）および各ウイルス株の LD

50

接種量

（FFU）

^小松川

1088

RABV-Dog

西ヶ原

CVS

ERA

1.0 10³ 

^NT

^NT

^NT

¹⁰⁰

(5/5)^a

¹⁰⁰

(5/5)

⁶⁰

(3/5)

1.0 10² 

¹⁰⁰ 

(10/10)

(10/10)¹⁰⁰ 

⁷⁰

(7/10)

⁸⁰

(4/5)

⁶⁰

(3/5)

¹⁰⁰

(5/5)

1.0 10¹ 

⁸⁰ 

(8/10)

⁵⁰

(5/10)

⁵⁰

(5/10)

⁴⁰

(2/5)

²⁰

(1/5)

²⁰

(1/5)

1.0 10⁰ 

⁰ 

(0/10)

⁰

(0/10)

¹⁰

(1/10)

⁰

(0/5)

⁰

(0/5)

⁰

(0/5)

LD50 (FFU)

4.2

10.0

15.3

20.8

48.1

ND

a  死亡した個体数  /  接種された個体数，  NT:  未実施，ND:  未決定 

   

                    

表 2-2.  小松川株，街上毒各株および固定毒各株を筋肉内接種されたマウスの          致死率（%）および各ウイルスの LD

50

接種量 

（FFU）

^小松川

¹⁰⁸⁸

^RABV-Dog

^西ヶ原

^CVS

^ERA

1.0 10⁶ 

^NT

^NT

^NT

¹⁰⁰

(5/5)^a

¹⁰⁰

(5/5)

⁴⁰

(2/5)

1.0 10⁵ 

^NT

^NT

^NT

⁴⁰

(2/5)

²⁰

(1/5)

²⁰

(1/5)

1.0 10⁴ 

^NT

¹⁰⁰

(10/10)

⁶⁰

(6/10)

⁰

(0/5)

⁰

(0/5)

⁰

(0/5)

1.0 10³ 

⁸⁰

(8/10)

²⁰

(2/10)

³⁰

(3/10)

^NT

^NT

^NT

1.0 10² 

²⁰

(2/10)

⁰

(0/10)

¹⁰

(1/10)

^NT

^NT

^NT

LD50 (FFU)

^{3.2 102}

^{2.4 103}

^{3.3 103}

^{1.5 105}

^{2.4 105}

^{>1.0 106}

a

 

死亡した個体数  /  接種された個体数  NT:  未実施

   

                    

   

図 2-3.  小松川株，街上毒各株および固定毒各株に感染したマウスの潜伏期間

3.0 LD50の各ウイルス株を１群 10 匹のマウス（6 週齢・雄）の左大腿筋に接種した後，接種日からマウ スの発症日までの日数（潜伏期間）を調べた。黒点がそれぞれのマウスの潜伏期間を示す。

a 

発症個体数  /  接種された個体数

   

CVS Nishigahara RABV Dog 1088

0 10 20 30 40 50

潜伏期間 (日)

平均潜伏期間

12.8 日

12.9 日 4.9 日 5.3 日 16.8 日

固定毒株 街上毒株

小松川

西ヶ原

    

図 2-4.  発症マウスの脳におけるウイルス感染細胞の分布

3.0 LD50の各ウイルス株をマウスの左大腿筋に接種した後，発症極期（小松川株:  感染後 14 日目，

1088 株: 14 日目，RABV-Dog 株: 14 日目，西ヶ原株: 6 日目，CVS 株: 7 日目）に採取した脳を用い て，抗 P 蛋白質抗体による免疫組織染色を実施した。なお，未感染については，ウイルスの希釈に使用し た培養液の接種後 35 日目の脳を用いた。

スケールバーは 3.5mm を示す。 

   

図2-4: 発症マウスの脳におけるウイルス感染細胞の分布

小松川 未感染

西ヶ原 CVS

固定毒株

1088 RABV-Dog

街上毒株