免疫组库数据分析||immunarch教程：克隆型分析

作者: 周运来就是我 | 来源:发表于2020-08-17 21:09 被阅读0次

immunarch — Fast and Seamless Exploration of Single-cell and Bulk T-cell/Antibody Immune Repertoires in R

Repertoire overlap and public clonotypes

免疫组重叠（Repertoire overlap）是最常用的度量Repertoire 相似度的方法。它是通过计算在给定的Repertoire 之间共享的克隆类型的特定统计量来实现的，也称为“公共”克隆类型。immunarch 提供了几个指标:-公共克隆型数量(.method = "public")) -一个经典的重叠相似性度量。

overlap coefficient (.method = "overlap") ：重叠相似度的标准化度量。它被定义为交集的大小除以两个集合中较小的部分。
Jaccard index (.method = "jaccard") ：它度量有限样本集之间的相似性，定义为交集的大小除以样本集并集的大小。
Tversky index (.method = "tversky") ：一种对集合的非对称相似度量，用来比较一个变体和一个原型。如果使用默认参数，它类似于Dice的系数。
cosine similarity (.method = "cosine") ：两个非零向量之间的相似性度量
Morisita’s overlap index (.method = "morisita") ：个体在总体中分散的统计方法。它用于比较样本之间的重叠。
incremental overlap ： overlaps of the N most abundant clonotypes with incrementally growing N (.method = "inc+METHOD", e.g., "inc+public" or "inc+morisita").

包含所描述方法的函数是repOverlap。同样，当输出被传递到vis()函数时，输出很容易被可视化，它完成了所有的工作:

library(immunarch)  # Load the package into R
data(immdata)  # Load the test dataset
imm_ov1 <- repOverlap(immdata$data, .method = "public", .verbose = F)
imm_ov2 <- repOverlap(immdata$data, .method = "morisita", .verbose = F)

p1 <- vis(imm_ov1)
p2 <- vis(imm_ov2, .text.size = 2)

p1 + p2

vis(imm_ov1, "heatmap2")

您可以轻松更改有效数字的数量:

p1 <- vis(imm_ov2, .text.size = 2.5, .signif.digits = 1)
p2 <- vis(imm_ov2, .text.size = 2, .signif.digits = 2)

p1 + p2

repOverlapAnalysis可以对计算得到的重叠测度函数进行分析。

# Apply different analysis algorithms to the matrix of public clonotypes:
# "mds" - Multi-dimensional Scaling
repOverlapAnalysis(imm_ov1, "mds")

Standard deviations (1, .., p=4):
[1] 0 0 0 0

Rotation (n x k) = (12 x 2):
               [,1]       [,2]
A2-i129 -18.7767715 -18.360817
A2-i131  29.9586985  -7.870441
A2-i133  28.1148594  22.629093
A2-i132 -44.3435640   6.221812
A4-i191  13.8586515   7.452149
A4-i192 -14.0065477  27.068830
MS1      -8.8469009  -8.151574
MS2      -0.9712073  -1.297017
MS3     -10.4398629   4.894354
MS4       0.5131505  10.471309
MS5      18.5153823 -12.628029
MS6       6.4241122 -30.429669


# "tsne" - t-Stochastic Neighbor Embedding
repOverlapAnalysis(imm_ov1, "tsne")


              DimI      DimII
A2-i129 -11.893405   70.95531
A2-i131 112.806943 -229.78268
A2-i133 -34.283164   47.07587
A2-i132 -44.726418   11.90656
A4-i191 -13.979182   10.05010
A4-i192 -13.316741   89.16606
MS1     -30.856320   78.41378
MS2     -32.951243   16.06630
MS3     -18.041903   75.90590
MS4     -24.965529   16.01290
MS5     120.335521 -229.87194
MS6      -8.128559   44.10184
attr(,"class")
[1] "immunr_tsne" "matrix"


# Visualise the results
repOverlapAnalysis(imm_ov1, "mds") %>% vis()

同样可以基于结果聚类。

# Clusterise the MDS resulting components using K-means
repOverlapAnalysis(imm_ov1, "mds+kmeans") %>% vis()

为了从repertoires 列表中构建一个包含所有clonotypes的庞大表，使用pubRep函数。

# Pass "nt" as the second parameter to build the public repertoire table using CDR3 nucleotide sequences
pr.nt <- pubRep(immdata$data, "nt", .verbose = F)
pr.nt

                                                  CDR3.nt Samples A2-i129 A2-i131 A2-i133 A2-i132
    1:                   TGCGCCAGCAGCTTGGAAGAGACCCAGTACTTC       8       1      NA       1       1
    2:                   TGTGCCAGCAGCTTCCAAGAGACCCAGTACTTC       7      NA       1       1       2
    3:                   TGTGCCAGCAGTTACCAAGAGACCCAGTACTTC       7       1       1       1      NA
    4:                   TGCGCCAGCAGCTTCCAAGAGACCCAGTACTTC       6       2      NA       1       1
    5:                      TGTGCCAGCAGCCAAGAGACCCAGTACTTC       6       4       2      NA       2
   ---                                                                                            
75101:             TGTGCTTCACAACTCTTATTGGACGAGACCCAGTACTTC       1      NA       1      NA      NA
75102: TGTGCTTCACAAGCCCTACAGGGCACTTTCCATAATTCACCCCTCCACTTT       1      NA      NA      NA      NA
75103:                   TGTGCTTCAGGGCGGGCCTACGAGCAGTACTTC       1      NA      NA      NA      NA
75104:             TGTGCTTCCGCCGGACCGGACCGGGAGACCCAGTACTTC       1      NA      NA       1      NA
75105:                TGTGCTTGCGGGACAGATAACTATGGCTACACCTTC       1      NA      NA      NA      NA
       A4-i191 A4-i192 MS1 MS2 MS3 MS4 MS5 MS6
    1:      NA       1  NA  NA   1   1   1   1
    2:       1      NA   1  NA  NA   2  NA   1
    3:       1       1   1  NA   2  NA  NA  NA
    4:      NA      NA  NA   1  NA   1  NA   1
    5:       3       1  NA  NA  NA  NA   4  NA
   ---                                        
75101:      NA      NA  NA  NA  NA  NA  NA  NA
75102:      NA      NA  NA  NA  NA  NA   1  NA
75103:      NA      NA   1  NA  NA  NA  NA  NA
75104:      NA      NA  NA  NA  NA  NA  NA  NA
75105:      NA       1  NA  NA  NA  NA  NA  NA

# Pass "aa+v" as the second parameter to build the public repertoire table using CDR3 aminoacid sequences and V alleles
# In order to use only CDR3 aminoacid sequences, just pass "aa"
pr.aav <- pubRep(immdata$data, "aa+v", .verbose = F)
pr.aav

                 CDR3.aa   V.name Samples A2-i129 A2-i131 A2-i133 A2-i132 A4-i191 A4-i192 MS1
    1:         CASSLEETQYF  TRBV5-1       8       1      NA       2       1      NA       2  NA
    2:     CASSDSSGGANEQFF  TRBV6-4       6       1       1       2      NA       3      NA  NA
    3:         CASSFQETQYF  TRBV5-1       6       3      NA       1       1      NA      NA  NA
    4:         CASSLGETQYF TRBV12-4       6       2      NA      NA       4       3      NA   1
    5:     CASSDSGGSYNEQFF  TRBV6-4       5      NA      NA      NA       3      NA       1   1
   ---                                                                                         
74440:     CTSSRPTQGAYEQYF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
74441:    CTSSSRAGAGTDTQYF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
74442: CTSSYPGLAGLKRKETQYF  TRBV7-2       1      NA      NA      NA       1      NA      NA  NA
74443:    CTSSYRQRPYQETQYF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
74444:      CTSSYSTSGVGQFF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
       MS2 MS3 MS4 MS5 MS6
    1:  NA   1   1   1   1
    2:  NA   2  NA  NA  12
    3:   1  NA   1  NA   1
    4:  NA  NA  NA   2   1
    5:  NA   1  NA  NA   1
   ---                    
74440:  NA  NA  NA  NA   1
74441:  NA   1  NA  NA  NA
74442:  NA  NA  NA  NA  NA
74443:  NA   1  NA  NA  NA
74444:  NA  NA   1  NA  NA

# You can also pass the ".coding" parameter to filter out all noncoding sequences first:
pr.aav.cod <- pubRep(immdata$data, "aa+v", .coding = T)
pr.aav.cod


                   CDR3.aa   V.name Samples A2-i129 A2-i131 A2-i133 A2-i132 A4-i191 A4-i192 MS1
    1:         CASSLEETQYF  TRBV5-1       8       1      NA       2       1      NA       2  NA
    2:     CASSDSSGGANEQFF  TRBV6-4       6       1       1       2      NA       3      NA  NA
    3:         CASSFQETQYF  TRBV5-1       6       3      NA       1       1      NA      NA  NA
    4:         CASSLGETQYF TRBV12-4       6       2      NA      NA       4       3      NA   1
    5:     CASSDSGGSYNEQFF  TRBV6-4       5      NA      NA      NA       3      NA       1   1
   ---                                                                                         
74440:     CTSSRPTQGAYEQYF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
74441:    CTSSSRAGAGTDTQYF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
74442: CTSSYPGLAGLKRKETQYF  TRBV7-2       1      NA      NA      NA       1      NA      NA  NA
74443:    CTSSYRQRPYQETQYF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
74444:      CTSSYSTSGVGQFF  TRBV7-2       1      NA      NA      NA      NA      NA      NA  NA
       MS2 MS3 MS4 MS5 MS6
    1:  NA   1   1   1   1
    2:  NA   2  NA  NA  12
    3:   1  NA   1  NA   1
    4:  NA  NA  NA   2   1
    5:  NA   1  NA  NA   1
   ---                    
74440:  NA  NA  NA  NA   1
74441:  NA   1  NA  NA  NA
74442:  NA  NA  NA  NA  NA
74443:  NA   1  NA  NA  NA
74444:  NA  NA   1  NA  NA


# Create a public repertoire with coding-only sequences using both CDR3 amino acid sequences and V genes
pr <- pubRep(immdata$data, "aa+v", .coding = T, .verbose = F)

# Apply the filter subroutine to leave clonotypes presented only in healthy individuals
pr1 <- pubRepFilter(pr, immdata$meta, c(Status = "C"))

# Apply the filter subroutine to leave clonotypes presented only in diseased individuals
pr2 <- pubRepFilter(pr, immdata$meta, c(Status = "MS"))

# Divide one by another
pr3 <- pubRepApply(pr1, pr2)

# Plot it
p <- ggplot() +
  geom_jitter(aes(x = "Treatment", y = Result), data = pr3)
p

参考：
https://immunarch.com/articles/web_only/v4_overlap.html

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本文标题：免疫组库数据分析||immunarch教程：克隆型分析

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免疫组库数据分析||immunarch教程：克隆型分析

Repertoire overlap and public clonotypes

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