Package 'dowser'

Description

airrClone defines a common data structure for perform lineage reconstruction from AIRR data, based heavily on alakazam::ChangeoClone.

Slots

data

data.frame containing sequences and annotations. Contains the columns sequence_id and sequence, as well as any additional sequence-specific annotation columns

clone

string defining the clone identifier

germline

string containing the heavy chain germline sequence for the clone

lgermline

string containing the light chain germline sequence for the clone

hlgermline

string containing the combined germline sequence for the clone

v_gene

string defining the V segment gene call

j_gene

string defining the J segment gene call

junc_len

numeric junction length (nucleotide count)

locus

index showing which locus represented at each site

region

index showing FWR/CDR region for each site

phylo_seq

sequence column used for phylogenetic tree building

numbers

index (usually IMGT) number of each site in phylo_seq

See Also

See formatClones for use.

Description

Same as ExampleClones but with biopsies predicted at internal nodes

Usage

BiopsyTrees

Format

A tibble of airrClone and phylo objects output by getTrees.

• clone_id: Clonal cluster

• data: List of airrClone objects

• seqs: Number of sequences

• trees: List of phylo objects

See Also

BiopsyTrees

Description

bootstrapTrees Phylogenetic bootstrap function.

Usage

bootstrapTrees(
  clones,
  bootstraps,
  nproc = 1,
  trait = NULL,
  dir = NULL,
  id = NULL,
  modelfile = NULL,
  build = "pratchet",
  exec = NULL,
  igphyml = NULL,
  fixtrees = FALSE,
  quiet = 0,
  rm_temp = TRUE,
  palette = NULL,
  resolve = 2,
  rep = NULL,
  keeptrees = TRUE,
  lfile = NULL,
  seq = NULL,
  downsample = FALSE,
  tip_switch = 20,
  boot_part = "locus",
  force_resolve = FALSE,
  ...
)

Arguments

Value

A list of trees and/or switch counts for each bootstrap replicate.

Description

Determine consensus clone sequence and create germline for clone

Usage

buildClonalGermline(
  receptors,
  references,
  chain = "IGH",
  use_regions = FALSE,
  vonly = FALSE,
  seq = "sequence_alignment",
  id = "sequence_id",
  clone = "clone_id",
  v_call = "v_call",
  j_call = "j_call",
  j_germ_length = "j_germline_length",
  j_germ_aa_length = "j_germline_aa_length",
  amino_acid = FALSE,
  ...
)

Arguments

Details

Return object adds/edits following columns:

• seq: Sequences potentially padded same length as germline

• germline_alignment: Full length germline

• germline_alignment_d_mask: Full length, D region masked

• vonly: V gene segment of germline if vonly=TRUE

• regions: String of VDJ segment in position if use_regions=TRUE

Value

Tibble with reconstructed germlines

See Also

createGermlines buildGermline, stitchVDJ

Description

Reconstruct germlines from alignment data.

Usage

buildGermline(
  receptor,
  references,
  seq = "sequence_alignment",
  id = "sequence_id",
  clone = "clone_id",
  v_call = "v_call",
  d_call = "d_call",
  j_call = "j_call",
  v_germ_start = "v_germline_start",
  v_germ_end = "v_germline_end",
  v_germ_length = "v_germline_length",
  d_germ_start = "d_germline_start",
  d_germ_end = "d_germline_end",
  d_germ_length = "d_germline_length",
  j_germ_start = "j_germline_start",
  j_germ_end = "j_germline_end",
  j_germ_length = "j_germline_length",
  np1_length = "np1_length",
  np2_length = "np2_length",
  amino_acid = FALSE
)

Arguments

Details

Return object contains multiple IMGT-gapped germlines:

• full: Full length germline

• dmask: Full length germline with D region masked

• vonly: V gene segment of germline

• regions: String showing VDJ segment of each position

Value

List of reconstructed germlines

See Also

buildClonalGermline, stitchVDJ

Description

Wrapper to build IgPhyML trees and infer intermediate nodes

Usage

buildIgphyml(
  clone,
  igphyml,
  trees = NULL,
  nproc = 1,
  temp_path = NULL,
  id = NULL,
  rseed = NULL,
  quiet = 0,
  rm_files = TRUE,
  rm_dir = NULL,
  partition = c("single", "cf", "hl", "hlf", "hlc", "hlcf"),
  omega = NULL,
  optimize = "lr",
  motifs = "FCH",
  hotness = "e,e,e,e,e,e",
  rates = NULL,
  asrc = 0.95,
  splitfreqs = FALSE,
  ...
)

Arguments

Details

Partition options in rate order:

• single: 1 omega for whole sequence

• cf: 2 omegas, 1 for all CDRs and 1 for all FWRs

• hl: 2 omegas, 1 for heavy and 1 for light chain

• hlf: 3 omegas, 1 for heavy FWR, 1 for all CDRs, and 1 for light FWRs

• hlc: 3 omegas, 1 for all FWRs, 1 for heavy CDRs, and 1 for light CDRs

• hlcf: 4 omegas, 1 for each heavy FWR, 1 for heavy CDR, 1 for light FWR, and 1 for light CDR

Value

phylo object created by igphyml with nodes attribute containing reconstructed sequences.

Description

Wrapper for alakazam::buildPhylipLineage

Usage

buildPhylo(
  clone,
  exec,
  temp_path = NULL,
  verbose = 0,
  rm_temp = TRUE,
  seq = "sequence",
  tree = NULL,
  onetree = TRUE
)

Arguments

Value

phylo object created by dnapars or dnaml with nodes attribute containing reconstructed sequences.

Description

Wrapper for phangorn::optim.pml

Usage

buildPML(
  clone,
  seq = "sequence",
  sub_model = "GTR",
  gamma = FALSE,
  asr = "seq",
  asr_thresh = 0.05,
  tree = NULL,
  data_type = "DNA",
  optNni = TRUE,
  optQ = TRUE,
  verbose = FALSE,
  resolve_random = TRUE,
  quiet = 0,
  rep = NULL
)

Arguments

Value

phylo object created by phangorn::optim.pml with nodes attribute containing reconstructed sequences.

Description

Wrapper for phangorn::pratchet

Usage

buildPratchet(
  clone,
  seq = "sequence",
  asr = "seq",
  asr_thresh = 0.05,
  tree = NULL,
  asr_type = "MPR",
  verbose = 0,
  resolve_random = TRUE,
  data_type = "DNA"
)

Arguments

Value

phylo object created by phangorn::pratchet with nodes attribute containing reconstructed sequences.

Description

Wrapper to build RAxML-ng trees and infer intermediate nodes

Usage

buildRAxML(
  clone,
  exec,
  seq = "sequence",
  sub_model = "GTR",
  partition = NULL,
  rseed = 28,
  name = "run",
  starting_tree = NULL,
  data_type = "DNA",
  from_getTrees = FALSE,
  rm_files = TRUE,
  asr = TRUE,
  rep = 1,
  dir = NULL,
  n_starts = NULL,
  ...
)

Arguments

Value

phylo object created by RAxML-ng with nodes attribute containing reconstructed sequences.

Description

calcRF Calculates the RF distance between two phylogenetic trees with the same tips and tip labels.

Usage

calcRF(tree_1, tree_2, nproc = 1)

Arguments

Value

The RF cluster value for the two input trees.

Description

collapseNodes Node collapsing function.

Usage

collapseNodes(trees, tips = FALSE, check = TRUE)

Arguments

Details

Use plotTrees(trees)[[1]] + geom_label(aes(label=node)) + geom_tippoint() to show node labels, and getSeq to return internal node sequences

Value

A tibble with phylo objects that have had internal nodes collapsed.

See Also

getTrees

Description

colorTree Gets a color palette for a predefined set of trait values

Usage

colorTrees(trees, palette, ambig = "blend")

Arguments

Details

Trees must have node states represented in a "states" vector. By default, ambiguous states (separated by ",") have their colors blended. If

Value

A list of colored trees

See Also

getPalette, getTrees, plotTrees

Description

condenseTrees Condenses a set of equally parsimonious node labels into a single tree

Usage

condenseTrees(trees, states, palette = NULL)

Arguments

Value

a phylo object representing all represented internal node states

Description

correlationTest performs root-to-tip regression date randomization test

Usage

correlationTest(
  clones,
  permutations = 1000,
  minlength = 0.001,
  perm_type = c("clustered", "uniform"),
  time = "time",
  sequence = "sequence_id",
  germline = "Germline",
  verbose = FALSE,
  polyresolve = TRUE,
  alternative = c("greater", "two.sided"),
  storeTree = FALSE,
  nproc = 1
)

Arguments

Details

Object returned contains these columns which are added or modified from input:

• data: airrClone object, same as input but with additional columns "cluster" which correspond to permutation cluster, and "divergence."

• slope: Slope of linear regression between divergence and time.

• correlation: Correlation between divergence and time.

• p: p value of correlation compared to permuted correlations.

• random_correlation: Mean correlation of permutation replicates.

• min_p: Minimum p value of data, determined by either the number of distinct clade/timepoint combinations or number of permutations.

• nposs: Number of possible distinct timepoint/clade combinations.

• nclust: Number of clusters used in permutation. If perm_type="uniform" this is the number of tips.

• p_gt/p_lt: P value that permuted correlations are greater or less than observed correlation. Only returned if alternative = "two.sided"

• test_trees: The phylo tree objects used, possibly with resolved polytomies.

Value

A tibble with the same columns as clones, but additional columns corresponding to test statistics for each clone.

See Also

Uses output from getTrees.

createGermlines Determine consensus clone sequence and create germline for clone

Description

createGermlines Determine consensus clone sequence and create germline for clone

Usage

createGermlines(
  data,
  references,
  locus = "locus",
  trim_lengths = FALSE,
  force_trim = FALSE,
  nproc = 1,
  seq = "sequence_alignment",
  v_call = "v_call",
  d_call = "d_call",
  j_call = "j_call",
  amino_acid = FALSE,
  id = "sequence_id",
  clone = "clone_id",
  v_germ_start = "v_germline_start",
  v_germ_end = "v_germline_end",
  v_germ_length = "v_germline_length",
  d_germ_start = "d_germline_start",
  d_germ_end = "d_germline_end",
  d_germ_length = "d_germline_length",
  j_germ_start = "j_germline_start",
  j_germ_end = "j_germline_end",
  j_germ_length = "j_germline_length",
  np1_length = "np1_length",
  np2_length = "np2_length",
  na.rm = TRUE,
  fields = NULL,
  verbose = 0,
  ...
)

Arguments

Details

Return object adds/edits following columns:

• seq: Sequences potentially padded same length as germline

• germline_alignment: Full length germline

• germline_alignment_d_mask: Full length, D region masked

• vonly: V gene segment of germline if vonly=TRUE

• regions: String of VDJ segment in position if use_regions=TRUE

Value

Tibble with reconstructed germlines

See Also

createGermlines buildGermline, stitchVDJ

Examples

vdj_dir <- system.file("extdata", "germlines", "imgt", "human", "vdj", package="dowser")
imgt <- readIMGT(vdj_dir)
db <- createGermlines(ExampleAirr[1,], imgt)

Description

Write a fasta file of sequences readFasta reads a fasta file

Usage

dfToFasta(
  df,
  file,
  id = "sequence_id",
  seq = "sequence",
  imgt_gaps = FALSE,
  columns = NULL
)

Arguments

Value

File of FASTA formatted sequences

Description

downsampleClone Down-sample clone to maximum tip/switch ratio

Usage

downsampleClone(clone, trait, tip_switch = 20, tree = NULL)

Arguments

Value

A vector with sequence for each locus at a specified node in tree.

Description

dowser is a phylogenetic analysis package as part of the Immcantation suite of tools. For additional details regarding the use of the dowser package see the vignettes:
browseVignettes("dowser")

References

1. Hoehn KB, Pybus OG, Kleinstein SH (2022) Phylogenetic analysis of migration, differentiation, and class switching in B cells. PLoS Computational Biology. https://doi.org/10.1371/journal.pcbi.1009885

Description

A small example database subset from Laserson and Vigneault et al, 2014.

Usage

ExampleAirr

Format

A data.frame with the following AIRR style columns:

• sequence_id: Sequence identifier

• sequence_alignment: IMGT-gapped observed sequence.

• germline_alignment_d_mask: IMGT-gapped germline sequence with N, P and D regions masked.

• v_call: V region allele assignments.

• v_call_genotyped: TIgGER corrected V region allele assignment.

• d_call: D region allele assignments.

• j_call: J region allele assignments.

• junction: Junction region sequence.

• junction_length: Length of the junction region in nucleotides.

• np1_length: Combined length of the N and P regions proximal to the V region.

• np2_length: Combined length of the N and P regions proximal to the J region.

• sample: Sample identifier. Time in relation to vaccination.

• isotype: Isotype assignment.

• duplicate_count: Copy count (number of duplicates) of the sequence.

• clone_id: Change-O assignment clonal group identifier.

References

1. Laserson U and Vigneault F, et al. High-resolution antibody dynamics of vaccine-induced immune responses. Proc Natl Acad Sci USA. 2014 111:4928-33.

See Also

ExampleDbChangeo ExampleClones

Description

A tibble of Ig lineage trees generated from the ExampleAirr file

Usage

ExampleClones

Format

A tibble of airrClone and phylo objects output by getTrees.

• clone_id: Clonal cluster

• data: List of airrClone objects

• seqs: Number of sequences

• trees: List of phylo objects

See Also

ExampleClones

Description

A small example database subset from Laserson and Vigneault et al, 2014.

Usage

ExampleDbChangeo

Format

A data.frame with the following Change-O style columns:

• SEQUENCE_ID: Sequence identifier

• SEQUENCE_IMGT: IMGT-gapped observed sequence.

• GERMLINE_IMGT_D_MASK: IMGT-gapped germline sequence with N, P and D regions masked.

• V_CALL: V region allele assignments.

• V_CALL_GENOTYPED: TIgGER corrected V region allele assignment.

• D_CALL: D region allele assignments.

• J_CALL: J region allele assignments.

• JUNCTION: Junction region sequence.

• JUNCTION_LENGTH: Length of the junction region in nucleotides.

• NP1_LENGTH: Combined length of the N and P regions proximal to the V region.

• NP2_LENGTH: Combined length of the N and P regions proximal to the J region.

• SAMPLE: Sample identifier. Time in relation to vaccination.

• ISOTYPE: Isotype assignment.

• DUPCOUNT: Copy count (number of duplicates) of the sequence.

• CLONE: Change-O assignment clonal group identifier.

References

1. Laserson U and Vigneault F, et al. High-resolution antibody dynamics of vaccine-induced immune responses. Proc Natl Acad Sci USA. 2014 111:4928-33.

See Also

ExampleAirr ExampleClones

Description

A small example database subset from Turner, J. S. et al. Human germinal centres engage memory and naive B cells after influenza vaccination. Nature 586, 127–132 (2020).

Usage

ExampleMixedClones

Format

A data.frame with the following Change-O style columns:

• clone_id: Clonal cluster

• data: List of airrClone objects

• locus: Locus identifier.

• seqs: Number of sequences

• igphyml_partitioned_trees: IgPhyML partitioned tree

• raxml_partitioned_trees: RAxML partitioned tree

Description

A small example database subset from Turner, J. S. et al. Human germinal centres engage memory and naive B cells after influenza vaccination. Nature 586, 127–132 (2020).

Usage

ExampleMixedDb

Format

A data.frame with the following Change-O style columns:

• sequence_id: Sequence identifier

• sequence: B cell sequence

• productive: A logical indicating if the sequence is productive.

• v_call: V region allele assignments.

• d_call: D region allele assignments.

• j_call: J region allele assignments.

• sequence_alignment: Sequence alignment.

• germline_alignment: Germline alignment without gaps.

• junction: Junction

• juncation_aa: Junction aa

• vj_inframe: A logical to see if the vj genes are in frame

• stop_codon: A indicator if there is a stop codon within the alignment

• locus: Locus identifier.

• v_sequence_start: Where the V gene starts

• v_sequence_end: Where the V gene ends

• v_germline_start: Where the V germline starts

• v_germline_end: Where the V germline ends

• np1_length: Length of np1

• d_sequence_start: Where the D gene starts

• d_sequence_end: Where the D gene ends

• d_germline_start: Where the D germline starts

• d_germline_end: Where the D germline ends

• np2_length: Length of np2

• j_sequence_start: Where the J gene starts

• j_sequence_end: Where the J gene ends

• j_germline_start: Where the J germline starts

• j_germline_end: Where the J germline ends

• junction_length: Length of the junction region in nucleotides.

• v_score: V score

• v_identity: Identity score of V

• v_support: V support

• d_score: D score

• d_identity: D identity

• d_support: D support

• j_score: J score

• j_support: J support

• j_identity: J identity

• cell_id: Cell identifier

• consensus_count: Consensus count

• indels: Logical if indels are present

• sequence_vdj: VDJ sequence

• v_germ_start_vdj: Where the V germline starts on the VDJ

• v_germ_end_vdj: Where the V germline ends on the VDJ

• subject: Subject identifier

• timepoint: Day the sample was taken

• cell_type: Type of cell

• replicate: Replicate number

• clone_id: Change-O assignment clonal group identifier.

• seq_type: Identifier of data type (10x)

• vj_gene: VJ gene

• vj_alt_gene: Alternative VJ gene

• v_germline_length: Length of the V germline segment

• d_germline_length: Length of the D germline segment

• j_germline_lenght: Length of the J germline segment

• germline_alignment_d_mask: Germline alignment with gaps

Description

exportTrees Exports phylogenetic trees

Usage

exportTrees(clones, filepath, tree_column = "trees", ...)

Arguments

Description

findSwitches Phylogenetic bootstrap function.

Usage

findSwitches(
  clones,
  permutations,
  trait,
  igphyml,
  fixtrees = FALSE,
  downsample = TRUE,
  tip_switch = 20,
  nproc = 1,
  dir = NULL,
  id = NULL,
  modelfile = NULL,
  build = "pratchet",
  exec = NULL,
  quiet = 0,
  rm_temp = TRUE,
  palette = NULL,
  resolve = 2,
  rep = NULL,
  keeptrees = FALSE,
  lfile = NULL,
  seq = NULL,
  boot_part = "locus",
  force_resolve = FALSE,
  ...
)

Arguments

Details

Tree building details are the same as getTrees. If keeptrees=TRUE (default) the returned object will contain a list named "trees" which contains a list of estimated tree objects for each bootstrap replicate. The object is structured like: trees[[<replicate>]][[<tree index>]]. If igphyml is specified (as well as trait), the returned object will contain a tibble named "switches" containing switch count information. This object can be passed to testSP and other functions to perform parsimony based trait value tests.

Trait values cannot contain values N, UCA, or NTIP. These are reserved for use by test statistic functions.

Value

A list of trees and/or switch counts for each bootstrap replicate.

See Also

Uses output from formatClones with similar arguments to getTrees. Output can be visualized with plotTrees, and tested with testPS, testSC, and testSP.

Examples

## Not run: 
data(ExampleAirr)
ExampleAirr$sample_id <- sample(ExampleAirr$sample_id)
clones <- formatClones(ExampleAirr, trait="sample_id")

igphyml <- "~/apps/igphyml/src/igphyml"
btrees <- findSwitches(clones[1:2,], permutations=10, nproc=1,
   igphyml=igphyml, trait="sample_id")
plotTrees(btrees$trees[[4]])[[1]]
testPS(btrees$switches)

## End(Not run)

Description

formatClones takes a data.frame or tibble with AIRR or Change-O style columns as input and masks gap positions, masks ragged ends, removes duplicates sequences, and merges annotations associated with duplicate sequences. If specified, it will un-merge duplicate sequences with different values specified in the traits option. It returns a list of airrClone objects ordered by number of sequences which serve as input for lineage reconstruction.

Usage

formatClones(
  data,
  seq = "sequence_alignment",
  clone = "clone_id",
  subgroup = "clone_subgroup",
  id = "sequence_id",
  germ = "germline_alignment_d_mask",
  v_call = "v_call",
  j_call = "j_call",
  junc_len = "junction_length",
  mask_char = "N",
  max_mask = 0,
  pad_end = TRUE,
  text_fields = NULL,
  num_fields = NULL,
  seq_fields = NULL,
  add_count = TRUE,
  verbose = FALSE,
  collapse = TRUE,
  cell = "cell_id",
  locus = "locus",
  traits = NULL,
  mod3 = TRUE,
  randomize = TRUE,
  use_regions = TRUE,
  dup_singles = FALSE,
  nproc = 1,
  chain = "H",
  heavy = "IGH",
  filterstop = TRUE,
  minseq = 2,
  split_light = FALSE,
  light_traits = FALSE,
  majoronly = FALSE,
  columns = NULL
)

Arguments

Details

This function is a wrapper for makeAirrClone. Also removes whitespace, ;, :, and = from ids

Value

A tibble of airrClone objects containing modified clones.

See Also

Executes in order makeAirrClone. Returns a tibble of airrClone objects which serve as input to getTrees and findSwitches.

Examples

data(ExampleAirr)
# Select two clones, for demonstration purpose
sel <- c("3170", "3184")
clones <- formatClones(ExampleAirr[ExampleAirr$clone_id %in% sel,],traits="sample_id")

Description

getNodeSeq Sequence retrieval function.

Usage

getAllSeqs(data, imgt_gaps = TRUE)

Arguments

Details

Column names: clone_id = clone id node_id = name of node, either the sequence name if a tip or Node<number> if internal node node = node number in tree. Tips are nodes 1:<number of tips>. locus = locus of sequence sequence = ungapped sequence, either observed for tips or reconstructed for internal nodes sequence_alignment = sequence with IMGT gaps (optional)

Value

A tibble with sequence information for each tip and internal node of a set of trees.

See Also

getTrees getNodeSeq

Description

getBootstraps Phylogenetic bootstrap function.

Usage

getBootstraps(
  clones,
  bootstraps,
  nproc = 1,
  bootstrap_nodes = TRUE,
  dir = NULL,
  id = NULL,
  build = "pratchet",
  exec = NULL,
  quiet = 0,
  rm_temp = TRUE,
  rep = NULL,
  seq = NULL,
  boot_part = "locus",
  by_codon = TRUE,
  starting_tree = FALSE,
  switches = FALSE,
  ...
)

Arguments

Value

The input clones tibble with an additional column for the bootstrap replicate trees.

Description

getDivergence get sum of branch lengths leading from the root of the tree. If the germline sequence is included in the tree, this will equal the germline divergence. If germline removed, this will equal the MRCA divergence

Usage

getDivergence(phy, minlength = 0.001)

Arguments

Value

A named vector of each tip's divergence from the tree's root.

getGermline get germline segment from specified receptor and segment

Description

getGermline get germline segment from specified receptor and segment

Usage

getGermline(
  receptor,
  references,
  segment,
  field,
  germ_start,
  germ_end,
  germ_length,
  germ_aa_start,
  germ_aa_length,
  amino_acid = FALSE
)

Arguments

Value

String of germline sequence from specified segment aligned with the sequence in the seq column of receptor.

Description

getNodeSeq Sequence retrieval function.

Usage

getNodeSeq(data, node, tree = NULL, clone = NULL, gaps = TRUE)

Arguments

Details

Use plotTrees(trees)[[1]] + geom_label(aes(label=node))+geom_tippoint() to show node labels, and getNodeSeq to return internal node sequences

Value

A vector with sequence for each locus at a specified node in tree.

See Also

getTrees

Description

getPalette Gets a color palette for a predefined set of trait values

Usage

getPalette(states, palette)

Arguments

Value

A named vector with each state corresponding to a color

See Also

getTrees, plotTrees

Description

getSeq Sequence retrieval function.

Usage

getSeq(data, node, tree = NULL, clone = NULL, gaps = TRUE)

Arguments

Value

A vector with sequence for each locus at a specified node in tree.

See Also

getTrees

Description

getSubClones plots a tree or group of trees

Usage

getSubclones(
  heavy,
  light,
  nproc = 1,
  minseq = 1,
  id = "sequence_id",
  seq = "sequence_alignment",
  clone = "clone_id",
  cell = "cell_id",
  v_call = "v_call",
  j_call = "j_call",
  junc_len = "junction_length",
  nolight = "missing"
)

Arguments

Value

a tibble containing

Description

getSubTaxa Gets the tip labels from a clade

Usage

getSubTaxa(node, tree)

Arguments

Value

A vector containing tip labels of the clade

Examples

# Get taxa from all subtrees
data(BiopsyTrees)
tree <- BiopsyTrees$trees[[8]]
all_subtrees <- lapply(1:length(tree$nodes), function(x)getSubTaxa(x, tree))

Description

getTrees Tree building function.

Usage

getTrees(
  clones,
  trait = NULL,
  id = NULL,
  dir = NULL,
  modelfile = NULL,
  build = "pratchet",
  exec = NULL,
  igphyml = NULL,
  fixtrees = FALSE,
  nproc = 1,
  quiet = 0,
  rm_temp = TRUE,
  palette = NULL,
  seq = NULL,
  collapse = FALSE,
  check_divergence = TRUE,
  ...
)

Arguments

Details

Estimates phylogenetic tree topologies and branch lengths for a list of airrClone objects. By default, it will use phangorn::pratchet to estimate maximum parsimony tree topologies, and ape::acctran to estimate branch lengths. If igpyhml is specified, internal node trait values will be predicted by maximum parsimony. In this case, dir will need to be specified as a temporary directory to place all the intermediate files (will be created if not available). Further, id will need to specified to serve as a unique identifier for the temporary files. This should be chosen to ensure that multiple getTrees calls using the same dir do not overwrite each others files.

modelfile is written automatically if not specified, but doesn't include any constraints. Intermediate files are deleted by default. This can be toggled using (rm_files).

For examples and vignettes, see https://dowser.readthedocs.io

Value

A list of phylo objects in the same order as data.

See Also

formatClones, findSwitches, buildPhylo, buildPratchet, buildPML, buildIgphyml, buildRAxML

Examples

data(ExampleClones)
trees <- getTrees(ExampleClones[10,])
plotTrees(trees)[[1]]

## Not run: 
data(ExampleClones)

trees <- getTrees(ExampleClones[10,],igphyml="/path/to/igphyml",
         id="temp",dir="temp", trait="sample_id")
plotTrees(trees)[[1]]

## End(Not run)

Description

Same as ExampleClones but with isotypes predicted at internal nodes

Usage

IsotypeTrees

Format

A tibble of airrClone and phylo objects output by getTrees.

• clone_id: Clonal cluster

• data: List of airrClone objects

• seqs: Number of sequences

• trees: List of phylo objects

See Also

IsotypeTrees

Description

makeAirrClone takes a data.frame with AIRR or Change-O style columns as input and masks gap positions, masks ragged ends, removes duplicates sequences, and merges annotations associated with duplicate sequences. It returns a airrClone object which serves as input for lineage reconstruction.

Usage

makeAirrClone(
  data,
  id = "sequence_id",
  seq = "sequence_alignment",
  germ = "germline_alignment_d_mask",
  v_call = "v_call",
  j_call = "j_call",
  junc_len = "junction_length",
  clone = "clone_id",
  subgroup = "clone_subgroup",
  mask_char = "N",
  max_mask = 0,
  pad_end = TRUE,
  text_fields = NULL,
  num_fields = NULL,
  seq_fields = NULL,
  add_count = TRUE,
  verbose = FALSE,
  collapse = TRUE,
  chain = "H",
  heavy = NULL,
  cell = "cell_id",
  locus = "locus",
  traits = NULL,
  mod3 = TRUE,
  randomize = TRUE,
  use_regions = TRUE,
  dup_singles = FALSE,
  light_traits = FALSE
)

Arguments

Details

The input data.frame (data) must columns for each of the required column name arguments: id, seq, germ, v_call, j_call, junc_len, and clone. Additional annotation columns specified in the traits, text_fields, num_fields or seq_fields arguments will be retained in the data slot of the return object, but are not required. These options differ by their behavior among collapsed sequences. Identical sequences that differ by any values specified in the traits option will be kept distinct. Identical sequences that differ only by values in the num_fields option will be collapsed and the values of their num_fields columns will be added together. Similar behavior occurs with text_fields but the unique values will concatenated with a comma.

The default columns are IMGT-gapped sequence columns, but this is not a requirement. However, all sequences (both observed and germline) must be multiple aligned using some scheme for both proper duplicate removal and lineage reconstruction.

The value for the germline sequence, V-segment gene call, J-segment gene call, junction length, and clone identifier are determined from the first entry in the germ, v_call, j_call, junc_len and clone columns, respectively. For any given clone, each value in these columns should be identical.

To allow for cases where heavy and light chains are used, this function returns three sequence columns for heavy chains (sequence), light chain (lsequence, empty if none available), and concatenated heavy+light chain (hlsequence). These contain sequences in alignment with germline, lgermline, and hlgermline slots, respectively. The sequence column used for build trees is specified in the phylo_seq slot. Importantly, this column is also the sequence column that also has uninformative columns removed by cleanAlignment. It is highly likely we will change this system to a single sequence and germline slot in the near future.

The airrClone object also contains vectors locus, region, and numbers, which contain the locus, IMGT region, and IMGT number for each position in the sequence column specified in phylo_seq. If IMGT-gapped sequences are not supplied, this will likely result in an error. Specify use_regions=FALSE if not using IMGT-gapped sequences

Value

A airrClone object containing the modified clone.

See Also

Returns an airrClone. See formatClones to generate an ordered list of airrClone objects.

Examples

data(ExampleAirr)
airr_clone <- makeAirrClone(ExampleAirr[ExampleAirr$clone_id=="3184",])

Description

makeModelFile Filler

Usage

makeModelFile(file, states, constraints = NULL, exceptions = NULL)

Arguments

Details

Currently the only option for constraints is "irrev", which forbids switches moving from left to right in the states vector.

Value

Name of model file

See Also

readModelFile, getTrees, findSwitches

Description

maskCodons Masks codons split by insertions

Usage

maskCodons(
  id,
  q,
  s,
  keep_alignment = FALSE,
  gap_opening = 5,
  gap_extension = 1,
  keep_insertions = FALSE,
  mask = TRUE
)

Arguments

Details

Performs global alignment of q and s, masks codons in s that are split by insertions (see example) masking_note notes codon positions in subject_alignment sequence that were masked, if found. subject_alignment contains subject sequence aligned to query (q) sequence query_alignment contains query sequence aligned to subject (q) sequence sequence_masked will be NA if frameshift or alignment error detected/

Value

A list with split codons masked, if found (sequence_masked).

See Also

maskSequences, Biostrings::pairwiseAlignment.

Examples

s = "ATCATCATC..."
q = "ATCTTTATCATC"
print(maskCodons(1,q,s,TRUE))

s <- "ATCATCATC..."
q <- "ATTTTCATCATC"
print(maskCodons("test",q,s,keep_alignment=TRUE,keep_insertions=TRUE))

Description

maskSequences Mask codons split by insertions in V gene

Usage

maskSequences(
  data,
  sequence_id = "sequence_id",
  sequence = "sequence",
  sequence_alignment = "sequence_alignment",
  v_sequence_start = "v_sequence_start",
  v_sequence_end = "v_sequence_end",
  v_germline_start = "v_germline_start",
  v_germline_end = "v_germline_end",
  junction_length = "junction_length",
  keep_alignment = FALSE,
  keep_insertions = FALSE,
  mask_codons = TRUE,
  mask_cdr3 = TRUE,
  nproc = 1
)

Arguments

Details

Performs global alignment of sequence and sequence_alignment, masking codons in sequence_alignment that are split by insertions (see examples) masking_note notes codon positions in subject_alignment sequence that were masked, if found. subject_alignment contains subject sequence aligned to query sequence (only if keep_alignment=TRUE) query_alignment contains query sequence aligned to subject sequence (only if keep_alignment=TRUE) sequence_masked will be NA if frameshift or alignment error detected. This will be noted insertions column will be returned if keep_insertions=TRUE, contains a comma-separated list of each <position in query alignment>-<sequence>. See example. in masking_note.

Value

A tibble with masked sequence in sequence_masked column, as well as other columns.

See Also

maskCodons, Biostrings::pairwiseAlignment.

Description

plotTrees plots a tree or group of trees

Usage

plotTrees(
  trees,
  nodes = FALSE,
  tips = NULL,
  tipsize = NULL,
  scale = 0.01,
  palette = "Dark2",
  base = FALSE,
  layout = "rectangular",
  node_nums = FALSE,
  tip_nums = FALSE,
  title = TRUE,
  labelsize = NULL,
  common_scale = FALSE,
  ambig = "grey",
  bootstrap_scores = FALSE,
  tip_palette = NULL,
  node_palette = NULL,
  guide_title = NULL
)

Arguments

Details

Function uses ggtree functions to plot tree topologies estimated by getTrees, and findSwitches. Object can be further modified with ggtree functions. Please check out https://bioconductor.org/packages/devel/bioc/vignettes/ggtree/inst/doc/ggtree.html and cite ggtree in addition to dowser if you use this function.

Value

a grob containing a tree plotted by ggtree.

See Also

getTrees, findSwitches

Examples

data(ExampleClones)
trees <- getTrees(ExampleClones[10,])
plotTrees(trees)[[1]]

Description

Read a fasta file into a list of sequences readFasta reads a fasta file

Usage

readFasta(file)

Arguments

Value

List of sequences

Description

Loads all reference germlines from an Immcantation-formatted IMGT database.

Usage

readIMGT(dir, quiet = FALSE)

Arguments

Details

Input directory must be formatted to Immcantation standard. See https://changeo.readthedocs.io/en/stable/examples/igblast.html for example of how to download.

Value

List of lists, leading to IMGT-gapped nucleotide sequences. Structure of object is list[[locus]][[segment]] locus refers to locus (e.g. IGH, IGK, TRA) segment refers to gene segment category (V, D, or J)

Examples

# vdj_dir contains a minimal example of reference germlines 
# (IGHV3-11*05, IGHD3-10*01 and IGHJ5*02)
# which are the gene assignments for ExampleDb[1,]
vdj_dir <- system.file("extdata", "germlines", "imgt", "human", "vdj", package="dowser")
imgt <- readIMGT(vdj_dir)

Description

Read in all trees from a lineages file

Usage

readLineages(
  file,
  states = NULL,
  palette = NULL,
  run_id = "",
  quiet = TRUE,
  append = NULL,
  format = "nexus",
  type = "jointpars"
)

Arguments

Value

A list of phylo objects from file.

Description

readModelFile Filler

Usage

readModelFile(file, useambig = FALSE)

Arguments

Value

A named vector containing the states of the model

See Also

makeModelFile, findSwitches, getTrees

Description

reconIgPhyML IgPhyML parsimony reconstruction function

Usage

reconIgPhyML(
  file,
  modelfile,
  id,
  igphyml = "igphyml",
  mode = "switches",
  type = "recon",
  nproc = 1,
  quiet = 0,
  rm_files = FALSE,
  rm_dir = NULL,
  states = NULL,
  palette = NULL,
  resolve = 2,
  rseed = NULL,
  force_resolve = FALSE,
  ...
)

Arguments

Value

Either a tibble of switch counts or a list of trees with internal nodes predicted by parsimony.

Description

Reroot phylogenetic tree to have its germline sequence at a zero-length branch to a node which is the direct ancestor of the tree's UCA. Assigns uca to be the ancestral node to the tree's germline sequence, as germid as the tree's germline sequence ID.

Usage

rerootTree(tree, germline, min = 0.001, verbose = 1)

Arguments

Value

phylo object rooted at the specified germline

Description

resolveLightChains resolve light chain V and J subgroups within a clone

Usage

resolveLightChains(
  data,
  nproc = 1,
  minseq = 1,
  locus = "locus",
  heavy = "IGH",
  id = "sequence_id",
  seq = "sequence_alignment",
  clone = "clone_id",
  cell = "cell_id",
  v_call = "v_call",
  j_call = "j_call",
  junc_len = "junction_length",
  nolight = "missing",
  pad_ends = TRUE
)

Arguments

Details

1. Make temporary array containing light chain clones 2. Enumerate all possible V, J, and junction length combinations 3. Determine which combination is the most frequent 4. Assign sequences with that combination to clone t 5. Copy those sequences to return array 6. Remove all cells with that combination from temp array 7. Repeat 1-6 until temporary array zero. If there is more than rearrangement with the same V/J in the same cell, pick the one with the highest non-ambiguous characters. Cells with missing light chains are grouped with their subgroup with the closest matching heavy chain (Hamming distance) then the largest and lowest index subgroup if ties are present.

Outputs of the function are 1. clone_subgroup which identifies the light chain VJ rearrangement that sequence belongs to within it's clone 2. clone_subgroup_id which combines the clone_id variable and the clone_subgroup variable by a "_". 3. vj_cell which combines the vj_gene and vj_alt_cell columns by a ",".

Value

a tibble containing the same data as inputting, but with the column clone_subgroup added. This column contains subgroups within clones that contain distinct light chain V and J genes, with at most one light chain per cell.

Description

Resolve polytomies to have the minimum number of single timepoint clades

Usage

resolvePolytomies(
  phy,
  clone,
  minlength = 0.001,
  time = "time",
  sequence = "sequence_id",
  germline = "Germline",
  verbose = FALSE
)

Arguments

Details

Iteratively identifies polytomies (clusters of < minlength branches), prunes each descendant branch, combines clades with the same timepoint before grouping them back together. Checks to make sure that the divergence of each tip is the same after resolution.

Value

A phylo tree object in which polytomies are resolved to have the minimum number of single timepoint clades.

See Also

Uses output from getTrees during correlationTest.

Description

runCorrelationTest performs root-to-tip regression permutation test

Usage

runCorrelationTest(
  phy,
  clone,
  permutations,
  minlength = 0.001,
  polyresolve = TRUE,
  permutation = c("clustered", "uniform"),
  time = "time",
  sequence = "sequence_id",
  germline = "Germline",
  verbose = TRUE,
  alternative = c("greater", "two.sided")
)

Arguments

Details

See correlationTest for details

Value

A list of statistics from running the permutation test.

See Also

correlationTest.

Description

scaleBranches Branch length scaling function.

Usage

scaleBranches(clones, edge_type = "mutations")

Arguments

Details

Uses clones$trees[[1]]$edge_type to determine how branches are currently scaled.

Value

A tibble with phylo objects that have had branch lengths rescaled as specified.

See Also

getTrees

stitchRegions Similar to stitchVDJ but with segment IDs instead of nucleotides

Description

stitchRegions Similar to stitchVDJ but with segment IDs instead of nucleotides

Usage

stitchRegions(
  receptor,
  v_seq,
  d_seq,
  j_seq,
  np1_length = "np1_length",
  np2_length = "np1_length",
  n1_length = "n1_length",
  p3v_length = "p3v_length",
  p5d_length = "p5d_length",
  p3d_length = "p3d_length",
  n2_length = "n2_length",
  p5j_length = "p5j_length",
  np1_aa_length = "np1_aa_length",
  np2_aa_length = "np2_aa_length",
  amino_acid = FALSE
)

Arguments

Value

Full length germline VDJ sequence with segment IDs instead of nucleotides.

See Also

stitchVDJ

stitchVDJ combines germline gene segments to a single string

Description

stitchVDJ combines germline gene segments to a single string

Usage

stitchVDJ(
  receptor,
  v_seq,
  d_seq,
  j_seq,
  np1_length = "np1_length",
  np2_length = "np2_length",
  np1_aa_length = "np1_aa_length",
  np2_aa_length = "np2_aa_length",
  amino_acid = FALSE
)

Arguments

Value

Full length germline VDJ sequence aligned with aligned with the sequence in the seq column of receptor.

Description

testPS performs a PS test

Usage

testPS(
  switches,
  bylineage = FALSE,
  pseudocount = 0,
  alternative = c("less", "two.sided", "greater")
)

Arguments

Details

Output data table columns: RECON = PS for observed data PERMUTE = PS for permuted data DELTA = RECON - PERMUTE PLT = p value for DELTA < 0 PGT = p value for DELTA < 0

• RECON: PS for observed data.

• PERMUTE: PS for permuted data.

• DELTA: RECON - PERMUTE.

• PLT: p value that DELTA < 0

• PGT: p value that DELTA > 0

• STAT: Statistic used (PS).

• REP: Bootstrap repetition.

• REPS: Total number of bootstrap repetition.

Value

A list containing a tibble with mean PS statistics, and another with PS statistics per repetition.

See Also

Uses output from findSwitches. Related to testSP and testSC.

Examples

## Not run: 
igphyml <- "~/apps/igphyml/src/igphyml"
data(ExampleAirr)
ExampleAirr$sample_id <- sample(ExampleAirr$sample_id)
clones <- formatClones(ExampleAirr, trait="sample_id")
btrees <- findSwitches(clones[1:2], bootstraps=10, nproc=1,
   igphyml=igphyml, trait="sample_id")
testPS(btrees$switches)

## End(Not run)

Description

testSC performs an SC test

Usage

testSC(
  switches,
  dropzeroes = TRUE,
  bylineage = FALSE,
  pseudocount = 0,
  from = NULL,
  to = NULL,
  permuteAll = FALSE,
  alternative = c("two.sided", "greater", "less")
)

Arguments

Details

Output data table columns: RECON = SC for observed data PERMUTE = SC for permuted data DELTA = RECON - PERMUTE PLT = p value for DELTA < 0 PGT = p value for DELTA < 0

• FROM: State going from.

• TO: State going to.

• RECON: SC for observed data.

• PERMUTE: SC for permuted data.

• DELTA: RECON - PERMUTE.

• PLT: p value that DELTA < 0

• PGT: p value that DELTA > 0

• STAT: Statistic used (SC).

• REP: Bootstrap repetition.

• REPS: Total number of bootstrap repetition.

Value

A list containing a tibble with mean SC statistics, and another with SC statistics per repetition.

See Also

Uses output from findSwitches. Related to testPS and testSP.

Examples

## Not run: 
igphyml <- "~/apps/igphyml/src/igphyml"
data(ExampleAirr)
ExampleAirr$sample_id = sample(ExampleAirr$sample_id)
clones = formatClones(ExampleAirr, trait="sample_id")
btrees = findSwitches(clones[1:2], bootstraps=100, nproc=1,
   igphyml=igphyml, trait="sample_id", id="temp", dir="temp")
testSC(btrees$switches)

## End(Not run)

Description

testSP performs an SP test

Usage

testSP(
  switches,
  permuteAll = FALSE,
  from = NULL,
  to = NULL,
  dropzeroes = TRUE,
  bylineage = FALSE,
  pseudocount = 0,
  alternative = c("greater", "two.sided", "less"),
  tip_switch = 20,
  exclude = FALSE
)

Arguments

Details

Output data table columns: RECON = SP for observed data PERMUTE = SP for permuted data DELTA = RECON - PERMUTE PLT = p value for DELTA < 0 PGT = p value for DELTA < 0

• FROM: State going from.

• TO: State going to.

• RECON: SP for observed data.

• PERMUTE: SP for permuted data.

• DELTA: RECON - PERMUTE.

• PLT: p value that DELTA < 0

• PGT: p value that DELTA > 0

• STAT: Statistic used (SP).

• REP: Bootstrap repetition.

• REPS: Total number of bootstrap repetition.

Value

A list containing a tibble with mean SP statistics, and another with SP statistics per repetition.

See Also

Uses output from findSwitches. Related to testPS and testSC.

Examples

## Not run: 
igphyml <- "~/apps/igphyml/src/igphyml"
data(ExampleAirr)
ExampleAirr$sample_id = sample(ExampleAirr$sample_id)
clones = formatClones(ExampleAirr, trait="sample_id")
btrees = findSwitches(clones[1:2], bootstraps=10, nproc=1,
   igphyml=igphyml, trait="sample_id")
testSP(btrees$switches)

## End(Not run)

Description

Same as ExampleClones but with timepoint as a trait value

Usage

TimeTrees

Format

A tibble of airrClone and phylo objects output by getTrees.

• clone_id: Clonal cluster

• data: List of airrClone objects

• seqs: Number of sequences

• trees: List of phylo objects

See Also

TimeTrees

Description

treesToPDF exports trees to a pdf in an orderly fashion

Usage

treesToPDF(plots, file, nrow = 2, ncol = 2, ...)

Arguments

Value

a PDF of tree plots

See Also

plotTrees

Examples

## Not run: 
data(ExampleClones)
trees <- getTrees(ExampleClones[10,])
plots <- plotTrees(trees)
treesToPDF(plots,"test.pdf",width=5,height=6)

## End(Not run)

Description

writeCloneSequences Exports the sequences used in tree building.

Usage

writeCloneSequences(clones, file)

Arguments

Description

Write lineage file for IgPhyML use

Usage

writeLineageFile(
  data,
  trees = NULL,
  dir = ".",
  id = "N",
  rep = NULL,
  trait = NULL,
  empty = TRUE,
  partition = "single",
  heavy = "IGH"
)

Arguments

Value

Name of created lineage file.