UsageΒΆ
Warning
Using a local IEDB installation is strongly recommended for larger datasets or when the making predictions for many alleles, epitope lengths, or prediction algorithms. More information on how to install IEDB locally can be found on the Installation page.
usage: pvacsplice run [-h] [-e1 CLASS_I_EPITOPE_LENGTH]
[-e2 CLASS_II_EPITOPE_LENGTH]
[--iedb-install-directory IEDB_INSTALL_DIRECTORY]
[-b BINDING_THRESHOLD]
[--percentile-threshold PERCENTILE_THRESHOLD]
[--allele-specific-binding-thresholds]
[--aggregate-inclusion-binding-threshold AGGREGATE_INCLUSION_BINDING_THRESHOLD]
[--aggregate-inclusion-count-limit AGGREGATE_INCLUSION_COUNT_LIMIT]
[-m {lowest,median}] [-r IEDB_RETRIES] [-k]
[-t N_THREADS] [--pass-only]
[--normal-sample-name NORMAL_SAMPLE_NAME]
[--normal-cov NORMAL_COV] [--tdna-cov TDNA_COV]
[--trna-cov TRNA_COV] [--normal-vaf NORMAL_VAF]
[--tdna-vaf TDNA_VAF] [--trna-vaf TRNA_VAF]
[--tumor-purity TUMOR_PURITY]
[--maximum-transcript-support-level {1,2,3,4,5}]
[--biotypes BIOTYPES] [--net-chop-method {cterm,20s}]
[--netmhc-stab]
[--net-chop-threshold NET_CHOP_THRESHOLD]
[--problematic-amino-acids PROBLEMATIC_AMINO_ACIDS]
[--run-reference-proteome-similarity]
[--blastp-path BLASTP_PATH]
[--blastp-db {refseq_select_prot,refseq_protein}]
[--peptide-fasta PEPTIDE_FASTA] [-a {sample_name}]
[-s FASTA_SIZE] [--exclude-NAs] [-j JUNCTION_SCORE]
[-v VARIANT_DISTANCE] [-g]
[--anchor-types [{A,D,NDA,DA,N} [{A,D,NDA,DA,N} ...]]]
[--expn-val EXPN_VAL]
input_file sample_name allele
{BigMHC_EL,BigMHC_IM,DeepImmuno,MHCflurry,MHCflurryEL,MHCnuggetsI,MHCnuggetsII,NNalign,NetMHC,NetMHCIIpan,NetMHCIIpanEL,NetMHCcons,NetMHCpan,NetMHCpanEL,PickPocket,SMM,SMMPMBEC,SMMalign,all,all_class_i,all_class_ii}
[{BigMHC_EL,BigMHC_IM,DeepImmuno,MHCflurry,MHCflurryEL,MHCnuggetsI,MHCnuggetsII,NNalign,NetMHC,NetMHCIIpan,NetMHCIIpanEL,NetMHCcons,NetMHCpan,NetMHCpanEL,PickPocket,SMM,SMMPMBEC,SMMalign,all,all_class_i,all_class_ii} ...]
output_dir annotated_vcf ref_fasta gtf_file
Run the pVACsplice pipeline
positional arguments:
input_file RegTools junctions output TSV file
sample_name The name of the sample being processed. This will be
used as a prefix for output files.
allele Name of the allele to use for epitope prediction.
Multiple alleles can be specified using a comma-
separated list. For a list of available alleles, use:
`pvacsplice valid_alleles`.
{BigMHC_EL,BigMHC_IM,DeepImmuno,MHCflurry,MHCflurryEL,MHCnuggetsI,MHCnuggetsII,NNalign,NetMHC,NetMHCIIpan,NetMHCIIpanEL,NetMHCcons,NetMHCpan,NetMHCpanEL,PickPocket,SMM,SMMPMBEC,SMMalign,all,all_class_i,all_class_ii}
The epitope prediction algorithms to use. Multiple
prediction algorithms can be specified, separated by
spaces.
output_dir The directory for writing all result files.
annotated_vcf A VEP-annotated single- or multi-sample VCF containing
genotype and transcript information.The VCF may be
gzipped (requires tabix index).
ref_fasta A reference FASTA file. Note: this input should be the
same as the RegTools vcf input.
gtf_file A reference GTF file. Note: this input should be the
same as the RegTools gtf input.
optional arguments:
-h, --help show this help message and exit
-e1 CLASS_I_EPITOPE_LENGTH, --class-i-epitope-length CLASS_I_EPITOPE_LENGTH
Length of MHC Class I subpeptides (neoepitopes) to
predict. Multiple epitope lengths can be specified
using a comma-separated list. Typical epitope lengths
vary between 8-15. Required for Class I prediction
algorithms. (default: [8, 9, 10, 11])
-e2 CLASS_II_EPITOPE_LENGTH, --class-ii-epitope-length CLASS_II_EPITOPE_LENGTH
Length of MHC Class II subpeptides (neoepitopes) to
predict. Multiple epitope lengths can be specified
using a comma-separated list. Typical epitope lengths
vary between 11-30. Required for Class II prediction
algorithms. (default: [12, 13, 14, 15, 16, 17, 18])
--iedb-install-directory IEDB_INSTALL_DIRECTORY
Directory that contains the local installation of IEDB
MHC I and/or MHC II. (default: None)
-b BINDING_THRESHOLD, --binding-threshold BINDING_THRESHOLD
Report only epitopes where the mutant allele has ic50
binding scores below this value. (default: 500)
--percentile-threshold PERCENTILE_THRESHOLD
Report only epitopes where the mutant allele has a
percentile rank below this value. (default: None)
--allele-specific-binding-thresholds
Use allele-specific binding thresholds. To print the
allele-specific binding thresholds run `pvacsplice
allele_specific_cutoffs`. If an allele does not have a
special threshold value, the `--binding-threshold`
value will be used. (default: False)
--aggregate-inclusion-binding-threshold AGGREGATE_INCLUSION_BINDING_THRESHOLD
Threshold for including epitopes when creating the
aggregate report (default: 5000)
--aggregate-inclusion-count-limit AGGREGATE_INCLUSION_COUNT_LIMIT
Limit neoantigen candidates included in the aggregate
report to only the best n candidates per variant.
(default: 15)
-m {lowest,median}, --top-score-metric {lowest,median}
The ic50 scoring metric to use when filtering epitopes
by binding-threshold or minimum fold change. lowest:
Use the best MT Score and Corresponding Fold Change
(i.e. the lowest MT ic50 binding score and
corresponding fold change of all chosen prediction
methods). median: Use the median MT Score and Median
Fold Change (i.e. the median MT ic50 binding score and
fold change of all chosen prediction methods).
(default: median)
-r IEDB_RETRIES, --iedb-retries IEDB_RETRIES
Number of retries when making requests to the IEDB
RESTful web interface. Must be less than or equal to
100. (default: 5)
-k, --keep-tmp-files Keep intermediate output files. This might be useful
for debugging purposes. (default: False)
-t N_THREADS, --n-threads N_THREADS
Number of threads to use for parallelizing peptide-MHC
binding prediction calls. (default: 1)
--pass-only Only process VCF entries with a PASS status. (default:
False)
--normal-sample-name NORMAL_SAMPLE_NAME
In a multi-sample VCF, the name of the matched normal
sample. (default: None)
--normal-cov NORMAL_COV
Normal Coverage Cutoff. Only sites above this read
depth cutoff will be considered. (default: 5)
--tdna-cov TDNA_COV Tumor DNA Coverage Cutoff. Only sites above this read
depth cutoff will be considered. (default: 10)
--trna-cov TRNA_COV Tumor RNA Coverage Cutoff. Only sites above this read
depth cutoff will be considered. (default: 10)
--normal-vaf NORMAL_VAF
Normal VAF Cutoff in decimal format. Only sites BELOW
this cutoff in normal will be considered. (default:
0.02)
--tdna-vaf TDNA_VAF Tumor DNA VAF Cutoff in decimal format. Only sites
above this cutoff will be considered. (default: 0.25)
--trna-vaf TRNA_VAF Tumor RNA VAF Cutoff in decimal format. Only sites
above this cutoff will be considered. (default: 0.25)
--tumor-purity TUMOR_PURITY
Value between 0 and 1 indicating the fraction of tumor
cells in the tumor sample. Information is used during
aggregate report creation for a simple estimation of
whether variants are subclonal or clonal based on VAF.
If not provided, purity is estimated directly from the
VAFs. (default: None)
--maximum-transcript-support-level {1,2,3,4,5}
The threshold to use for filtering epitopes on the
Ensembl transcript support level (TSL). Keep all
epitopes with a transcript support level <= to this
cutoff. (default: 1)
--biotypes BIOTYPES A list of biotypes to use for pre-filtering
transcripts for processing in the pipeline. (default:
['protein_coding'])
--net-chop-method {cterm,20s}
NetChop prediction method to use ("cterm" for C term
3.0, "20s" for 20S 3.0). C-term 3.0 is trained with
publicly available MHC class I ligands and the authors
believe that is performs best in predicting the
boundaries of CTL epitopes. 20S is trained with in
vitro degradation data. (default: None)
--netmhc-stab Run NetMHCStabPan after all filtering and add
stability predictions to predicted epitopes. (default:
False)
--net-chop-threshold NET_CHOP_THRESHOLD
NetChop prediction threshold (increasing the threshold
results in better specificity, but worse sensitivity).
(default: 0.5)
--problematic-amino-acids PROBLEMATIC_AMINO_ACIDS
A list of amino acids to consider as problematic. Each
entry can be specified in the following format:
`amino_acid(s)`: One or more one-letter amino acid
codes. Any occurrence of this amino acid string,
regardless of the position in the epitope, is
problematic. When specifying more than one amino acid,
they will need to occur together in the specified
order. `amino_acid:position`: A one letter amino acid
code, followed by a colon separator, followed by a
positive integer position (one-based). The occurrence
of this amino acid at the position specified is
problematic., E.g. G:2 would check for a Glycine at
the second position of the epitope. The N-terminus is
defined as position 1. `amino_acid:-position`: A one
letter amino acid code, followed by a colon separator,
followed by a negative integer position. The
occurrence of this amino acid at the specified
position from the end of the epitope is problematic.
E.g., G:-3 would check for a Glycine at the third
position from the end of the epitope. The C-terminus
is defined as position -1. (default: None)
--run-reference-proteome-similarity
Blast peptides against the reference proteome.
(default: False)
--blastp-path BLASTP_PATH
Blastp installation path. (default: None)
--blastp-db {refseq_select_prot,refseq_protein}
The blastp database to use. (default:
refseq_select_prot)
--peptide-fasta PEPTIDE_FASTA
When running the reference proteome similarity step,
use this reference peptide FASTA file to find matches
instead of blastp. (default: None)
-a {sample_name}, --additional-report-columns {sample_name}
Additional columns to output in the final report. If
sample_name is chosen, this will add a column with the
sample name in every row of the output. This can be
useful if you later want to concatenate results from
multiple individuals into a single file. (default:
None)
-s FASTA_SIZE, --fasta-size FASTA_SIZE
Number of FASTA entries per IEDB request. For some
resource-intensive prediction algorithms like
Pickpocket and NetMHCpan it might be helpful to reduce
this number. Needs to be an even number. (default:
200)
--exclude-NAs Exclude NA values from the filtered output. (default:
False)
-j JUNCTION_SCORE, --junction-score JUNCTION_SCORE
Junction Coverage Cutoff. Only sites above this read
depth cutoff will be considered. (default: 10)
-v VARIANT_DISTANCE, --variant-distance VARIANT_DISTANCE
Regulatory variants can lie inside or outside of
splicing junction.Maximum distance window (upstream
and downstream) for a variant outside the junction.
(default: 100)
-g, --save-gtf Save a tsv file from the uploaded filtered GTF
data.Use this option to bypass GTF data upload time
for multiple pVACsplice runs. (default: False)
--anchor-types [{A,D,NDA,DA,N} [{A,D,NDA,DA,N} ...]]
The anchor types of junctions to use. Multiple anchors
can be specified using a comma-separated list.Choices:
A, D, NDA, DA, N (default: ['A', 'D', 'NDA'])
--expn-val EXPN_VAL Gene Expression cutoff. Only sites above this cutoff
will be considered. (default: 1.0)